METHOD AND DIAGNOSTIC KIT FOR DIAGNOSIS OF ENDOMETRIOSIS

Abstract: The invention relates to a method and a kit for the diagnosis of endometriosis using blood and endometrial leukocyte markers or a combination thereof. The marker is a surface antigen from endometrial or blood leukocytes.

Description: METHOD AND DIAGNOSTIC KIT FOR DIAGNOSIS OF ENDOMETRIOSIS BACKGROUND OF THE INVENTION (a) Field of the Invention The invention relates to a method and a kit for the diagnosis of endometriosis using blood and endometrial leukocyte markers, (b) Description of Prior Art Endometriosis is one of the most common gynecological disorders, affecting up to 15% of women within reproductive age. It is closely associated with severe pelvic pain, dysmenorrhea , dyspareunia, infertility and several other symptoms such as intraperitoneal bleeding, back pain, constipation and/or diarrhea. It is a major threat to physical, psychological and social integrity of the patients. Endometriosis is characterized by the implantation and growth of endometrial cells (which normally constitute the lining of the uterus) in extra-uterine sites such as the peritoneal cavity. Although the etiology and pathogenesis of endometriosis remain mainly unclear, the theory of retrograde menstruation is the most widely accepted to explain the presence of ectopic endometrial cells in the peritoneal cavity. However, this phenomenon occurs in most women and, thus, several other factors must be invoked to explain the implantation of endometrial cells and the subsequent development of endometriotic lesions. It is generally believed that initiation of endometriosis implies a complex cascade of events requiring several essential features . Retrogradely seeded endometrial cells must remain viable, be capable of adhering to the mesothelium and of proliferating. Local degradation of the extracellular matrix, as well as extensive vascularization, are also believed to play an essential role in promoting the invasion of the peritoneal cavity by endometrial cells. Furthermore, once implanted, ectopic endometrial cells must have the capacity to counteract the cytolytic action of the immune system. Indeed, this is supported by the observation of several immunological abnormalities in patients with endometriosis . At present, direct visualization of the endometriotic lesions under surgical procedures (laparascopy or laparotomy) is the golden standard and the only reliable method available to diagnose endometriosis. However, this method is highly invasive (i.e. surgery under general anesthesia), costly (i.e. direct cost and indirect cost due to convalescence) and requires a well -trained surgeon who has the ability to identify endometriotic lesions with a variety of appearances. The type of lesions, their size and their localization will determine the stage of the disease (stage I minimal, stage II mild, stage III moderate, stage IV severe) . However, there is still no clear consensus on how these parameters correlate with the stage of the disease and the prognostic of endometriosis. In addition, early or minimal endometriosis (which can involve microlesions) can be hardly diagnosed by surgical methods, as they are unlikely to be detected by direct visualization. Indeed, several studies have reported microscopic endometriotic lesions that were not detected laparoscopically . Because the diagnosis of endometriosis by surgical procedures is difficult, costly and invasive, in some cases, several physicians and patients tend to avoid it or at least seriously delay it. Hence, the length of time between the onset of symptoms and the diagnosis can be as long as 8 to 12 years. The possibility to diagnose endometriosis at an early stage would certainly improve the efficacy of the treatments, and reduce dramatically the number of years during which patients endure acute or chronic pain. Imaging methods such as transvaginal ultrasound and magnetic resonance imaging have been designed for the diagnosis of endometriosis. However, these techniques can only be reliable for the detection of large (> 1 cm diameter) endometriomas lesions detected among a very small proportion of patients with endometriosis. Moreover, the high cost of these techniques has limited their use for the diagnosis of endometriosis . Serum proteins such as CA-125 and placental protein-14 have been proposed as diagnostic markers for endometriosis. Elevated levels of CA-125 have been observed in serum, menstrual effluent and peritoneal fluid of patients with endometriosis. However, these markers, when used alone, are of very limited value for a diagnosis test. Indeed, these markers are not suitable for screening or diagnostic purposes because they provide poor sensitivity. Furthermore, levels of CA-125 and placental protein-14 vary according to several factors such as the assay, the stage of the disease and the menstrual cycle. Finally these markers are known to be modulated by conditions other than endometriosis . High concentrations of antibodies to endometrial antigens were found in the serum of patients with endometriosis, and thus were proposed as markers for a diagnostic test (International patent application publications WO 94/28021 and WO 92/18535) . However, the levels of specificity and sensibility with these tests remain very low. In most cases, the antigens recognized by these antibodies are still poorly characterized or yet totally unknown. In U.S. patent No. 5,478,725, low levels of αvβ3 integrin expression in endometrial samples during the secretory phase of the menstrual cycle is described as a predictor of endometriosis in infertile but not in fertile patients with endometriosis. This observation was associated with milder form of endometriosis (stages I and II) only and, thus, is not useful to detect advanced stages of the disease. Moreover, this method yielded a specificity of 91% but a very low sensitivity (38%) . Taking into account that a number immunological abnormalities have been reported in patients with endometriosis, it is conceivable that the proportion of leukocyte populations and/or their activation status may be modulated during the course of the disease and, thus, may provide some diagnostic value. Previous flow cytometric studies have shown that some T lymphocyte subpopulations (CD8+, CD45+/HLADR+, CD45+/CD3+/HLADR+ or CD3+/CD25+) can be slightly modulated in the peritoneal fluid of subjects with endometriosis relative to normal controls (Oosterlyncck D.J., et al . , Am J reprod. Immunol . , 31: 25-31, 1994; Becker J.L., et al . , Am J Reprod . Immunol . , 34: 179-187, 1995; Wu M.Y., et al . , Am. j. Reprod . Immunol . 35: 510-516, 1996). However, these observations have limited value for the diagnosis of endometriosis because peritoneal fluid collection is an invasive, non-conventional procedure. Proportions of leukocyte populations have also been studied in peripheral blood and endometrium of patients with endometriosis. Wu et al . , (supra) have reported a modest but significant decrease in the proportion of CD3+ T lymphocytes expressing either CD69 or CD25 activation marker in the blood of patients with advanced endometriosis but not in patients with mild stage of endometriosis or normal controls. This difference was observed in advanced cases of endometriosis only and it was too modest to be used as a diagnostic marker. In contrast, Oosterlynck et al . , (Oosterlyncck D.J., et al . , Am J " reprod . Immunol . , 31: 25-31, 1994) and Ho et al . (Ho H.N. , et al . , Hum Reprod. , 97: 2528-2533, 1997) reported no significant difference in term of T lymphocyte subpopulations when comparing endometriosis subjects with normal controls. These inconsistent results may be explained by the very low number of samples tested in these studies. Several studies have investigated whether leukocytes are also modulated in eutopic endometrium from patients with endometriosis. Results arising from these studies are contradictory, probably due to the fact that in most cases the methods used were only semi -quantitative and the number of samples tested were very low. For instance, by means of immunohistochemistry, Ota et al . (Ota H. , et al . , Am J Reprod . Immunol . , 35: 477-482, 1996) have reported that the number of CD3+ , CD4+ , or CD8+ T lymphocytes, cells bearing adhesion molecules (i.e. ICAM-1, LFA-1, CD2) or CD68+ cells were upregulated in the endometrium of patients with endometriosis compared with infertile controls. In contrast, several other studies using similar techniques have reported no difference in the proportion of T lymphocyte subsets (Klentzeris L.D., et al . , Eur. J Obstet gynecol Reprod Biol . , 63:41-47, 1995; Jones R.K., et al . , Fertil Steril , 66:81-89, 1996). In addition, a decrease in CD3 positive T cells has been shown by flow cytometry analysis but no difference in the proportion of CD4+ , CD8+ stromal leukocytes in the endometrium of patients with endometriosis compared with fertile controls. When these observations are tentatively used in a diagnostic test, they give only low levels of sensibility and specificity because of a significant overlap between the groups. Therefore, the diagnostic methods presented in the literature so far do not solve the problems encountered with the diagnosis of endometriosis by surgical procedures. It thus remains imperative to be provided with a less invasive, cheaper and reliable method that could allow detection of females suffering from endometriosis as early as possible. SUMMARY OF THE INVENTION One aim of the present invention is to provide a less invasive, cheaper and reliable method that could allow detection of females suffering from endometriosis as early as possible. In accordance with the present invention there is provided a specific blood and/or endometrial leukocyte marker for endometriosis selected from the group consisting of CD3+, CD4+, CD5+, CD8+, CD13+, CD14+, CD20+, CD36+, CD44+, CD56+, CD57+, CD69+, CD122+, HLADR+, CD16+, CD45RA+, CD45RO+, CD56-CD122+, CD3+CD4-CD69+, CD3 -CD8+HLADR- , CD3+CD4+, CD3+CD4-, CD3-CD4-, CD3+CD5+, CD3-CD5+, CD3-CD5-, CD3+CD8+, CD3+CD8-, CD3-CD8-, CD3+CD16+, CD3-CD16+, CD3+CD16-, CD3+CD20-, CD3-CD20-, CD3+CD44-, CD3-CD44+, CD3-CD44-, CD3+CD45RA-, CD3-CD45RA-, CD3+CD45RA+, CD3+CD45RO+, CD3-CD45RO+, CD3+CD45RO-, CD3+CD56-, CD3-CD56-, CD3+CD57-, CD3-CD57+, CD3-CD57-, CD3+CD69-, CD3+CD69+, CD3-CD69+, CD3+CD122-, CD3+HLADR+, CD3+HLADR-, CD3- HLADR+, CD3-HLADR-, CD4+CD13+, CD4+CD13-, CD4-CD13+, CD4+CD14-, CD4-CD14-, CD4-CD16-, CD4-CD36+, CD4+CD45RA-, CD4-CD45RA-, CD4+CD45RO+ , CD4+CD45RO-, CD4-CD45RO+, CD4+CD69-, CD4-CD69+, CD4-CD69-, CD4+HLADR-, CD4-HLADR+, CD8-CD44+, CD8-CD44-, CD8+CD69-, CD8+HLADR-, CD8-HLADR-, CD13+CD16-, CD13- CD16+, CD13+CD44+, CD13-CD44-, CD13+CD45RO- , CD13- CD45RO+, CD13-CD69+, CD13-CD122+, CD13-CD122-, CD13+HLADR+, CD13-HLADR+, CD14+CD13+, CD14+CD13-, CD14+CD16-, CD14+CD44+, CD14-CD44-, CD14+CD45RO+, CD14-CD69-, CD14+CD122-, CD14+HLADR+, CD14-HLADR+, CD20-CD5+, CD20-CD5-, CD20+CD44-, CD20-CD44+, CD20- CD44-, CD20-CD69+, CD20-CD69-, CD20+HLADR+, CD20- HLADR+, CD20-HLADR-, CD36-HLADR+ , CD56-CD16+, CD56- CD16-, CD56-CD44-, CD56+CD69-, CD56-CD69+, CD56-CD69-, CD56+CD122+, CD56+CD122-, CD56-CD122-, CD56+HLADR+, CD57-CD44+, CD57-CD44-, CD3 -CD4 -CD44+ , CDS- CD4+CD45RA-, CD3 -CD4-CD45RA+, CD3 -CD4-CD45RA- , CD3 - CD4-CD45RO+, CD3 -CD8-CD44+ , CD3+CD8+CD69- , CD3+CD8+HLADR-, CD3+CD8-HLADR+, CD3+CD8-HLADR- , CD3 - CD8-HLADR-, CD3+CD20-CD5+ , CD3+CD20-CD5- , CD3-CD20- CD5-, CD3+CD56+CD16+, CD3+CD56-CD16+ , CD3+CD56-CD16- , CD3-CD56+CD16+, CD3 -CD56+CD16- , CD3+CD56-CD44+, CD3+CD56-CD44-, CD3+CD56-CD122+ , CD3+CD56-CD122- , CD3 - CD56+CD122+, CD3 -CD56-CD122- , CD3 -CD56-HLADR- , CD3- CD57+CD44-, CD3 -CD57-CD44+ , CD3 -CD57-CD44- , CD3+CD57- HLADR+, CD4-CD13+CD16+, CD4-CD13 -CD16+ , CD4 -CD13 -CD16- , CD14+CD13+CD16b+, CD14+CD13+CD16b- , CD14+CD13 -CD16b- , CD14-CD13-HLADR+, CD14-CD13 -HLADR- , CD14+CD20+CD44+, CD14+CD20+CD44-, CD14+CD20-CD44+ , CD14+CD20-CD44- , CD14-CD20+CD44-, ratio CD3/CD45RO, Ratio CD13/CD3, Ratio CD13/CD8, Ratio CD14/CD3, and Ratio CD14/CD8. Also in accordance with the present invention, there is provided a diagnostic method for the detection of endometriosis in a patient sample. The method comprises the step of detecting at least one specific marker as described above, whereby detection of this specific marker is indicative of endometriosis . Further in accordance with the present invention, there is provided a diagnostic method for the detection of endometriosis in a patient sample. The method comprises the step of detecting at least two different surface antigens from blood or endometrial leukocytes, whereby detection of at least two different surface antigens is indicative of endometriosis. In accordance with a further embodiment of the invention, there is provided a diagnostic method for the detection of endometriosis in a patient sample. The method comprises the step of detecting a specific marker combination for endometriosis as defined above, whereby detection of this combination is indicative of endometriosis . Further in accordance with the present invention, there is provided a diagnostic kit for the detection of endometriosis. The kit comprises an antibody specific for the specific maker as described above. Preferably, the kit comprises at least two different antibodies, each specific for different surface antigens as defined in the specific marker combination defined above. Most preferably, the specific marker combination of the diagnostic kit is selected from the combination described below in Tables 1 and 2. For the purpose of the present invention, the following symbol "/" is intended to mean a ratio between an expression in front of the symbol and another expression after the symbol . BRIEF DESCRIPTION OF THE DRAWING Fig. 1 illustrates a predictive algorithm for the diagnosis of endometriosis. DETAILED DESCRIPTION OF THE INVENTION In accordance with the present invention, there is provided reliable diagnostic test for endometriosis that is less invasive and less costly than the actual surgical procedure accepted as the golden standard. An extensive study was undertaken by means of flow cytometric analysis, in which the proportion of several blood and endometrial leukocyte subsets was compared in patients with endometriosis and normal controls . The present invention identifies a series of leukocyte subsets that can be used as markers in a diagnostic test for endometriosis. These leukocyte subsets are defined according to the expression of cell surface antigens. Several cell surface antigens may define the same population of cells, and thus they are included in the present invention. Any other antibodies or molecules recognizing the same antigen or a different epitope, isoform, subunit, chain, glycosylation or phosphorylation form or an allelic variant of the same antigen, a member of the same complex, or an antigen with the same cell distribution is also included in the present invention. Further in accordance with the present invention, there is provided examples showing how at least two different surface antigens from blood and/or endometrial leukocytes can be used in combinations in a diagnostic test for endometriosis (Tables 1 and 2) . TABLE 1 Levels of sensibility and specificity provided by several examples of endometrial and/or blood marker combinations used as a diagnostic method for endometriosis Marker combination Threshold Specificity Sensibility value 1 Endometrial leukocyte markers 1. CD4+(<17%) 2 >67% 90% 67% CD8+CD69-(<21%) CD13+CD45RO- (< 17.5%) 2. CD4+(<15.5%) >60% 89% 65% CD8+CD69-(<21%) CD56+CD122-(>19%) CD3+CD45RA- (<35%) CD13+CD45RO- (< 17.5%) 3. CD4+(<17%) >67% 88% 65% CD8+CD69-(<21%) CD13-CD122+(>28%) CD13+CD45RO- (<17.5%) CD4+(<17%) >67% 89% 63% CD8+CD69-(<21%) CD14+CD13-CD16b- (>14.5%) CD3+CD16-(<40%) >55% 84% 62% CD13+CD45RO- (<13.5%) CD3+ (<40%) CD8+ (<20%) CD3+CD69+(<15%) 6. CD3+ (<40%) >65% 84% 63% CD3+CD8+(<16%) CD13+CD45RO- (<17.5%) CD3-CD20- (>56%) CD3+CD8+(<16%) >65% 81% 65% CD13+CD45RO- (<17.5%) CD3+CD5+ (<37%) CD3+CD122-(<42.5%) CD3-CD20- (>56%) CD3+CD45RO- (<30%) Marker combination Threshold Specificity Sensibility value 1 8. CD3+CD8+(<16%) >60% 82% 64% CD13+CD45RO-(<17.5%) CD3+CD5+ (<37%) CD3+CD122-(<42.5%) CD3+CD20-CD5- (>7.7%) >60% 81% 66% CD4+CD13-(<20.5%) CD56-CD122-(<47%) 10. CD3+CD8+ (<16%) >60% 80% 65% CD13+CD45RO-(<17.5%) CD4+CD45RA-(<16%) CD3+CD45RO- (<30%) 11. CD3+ (<40%) CD8+CD69-(<18%) CD3-CD4-CD45RO+ (>56%) Ratio CD13+/CD3+ (>0.675%) CD13+CD45RO-(<21%) 12. CD3+CD8+(<16%) >70% 81% 61% CD13+CD45RO- (<17.5%) CD3-CD5- (>54%) CD20-CD5+ (<44%) 13. CD8+ (<20%) 2 >51% 81% 60% CD5+ (<37%) CD3-CD20- (>58%) CD3-HLADR- (>54.5%) 14. CD3+CD8+ (<16%) >60% 81% 60% CD13+CD45RO- (<17.5%) CD5+ (<40%) 15. CD4+(<17%) >50% 76% 71% CD13-CD122+(>28%) CD8+CD69-(<19.5%) CD3+CD45RA- (<37%) Marker combination Threshold Specificity Sensibility value 1 16. CD4+(<15.5%) >35% 71% 78% CD8+CD69-(<21%) CD13-CD122+(>28%) CD3+CD45RA-(<35%) CD13+CD45RO- (<17.5%) 17. CD4+(<15.5%) >40% 70% 78% CD8+CD69-(<21%) CD56-CD122-(<47%) CD3+CD45RA- (<35%) CD13+CD45RO- (<17.5%) 18. CD3+(<40%) >35% 72% 76% CD4+(<17%) CD3+CD8+(<16%) CD13+CD45RO-(<21%) CD3+CD5+ (<37%) 19. CD3+(<40%) >40% 74% 74% CD4+(<17%) CD3+CD8+(<16%) CD13+CD45RO-(<21%) 20. CD3+(<40%) >40% 75% 73% CD3+CD8+(<16%) CD13+CD45RO-(<21%) 21. CD3+ (<40%) >25% 71% 69% CD5+ (<40%) CD3+CD5+ (<37%) CD69+ (<33%) CD4-CD69+ (<35%) 22. CD3+ (<40%) >30% 68% 83% CD3+CD8+(<13.5%) CD13+CD45RO- (<17.5%) 23. CD3+ (<40%) >30% 61% 86% CD3+CD8+(<16%) CD13+CD45RO- (<17.5%) Marker combination Threshold Specificity Sensibility value 1 24. CD3+ (<40%) >22% 62% 80% CD3+CD8+ (<16%) CD13+CD45RO- (<17.5%) CD3-CD20- (>58%) CD56-CD16- (<46%) 25. CD3+CD16- (<47.5%) >45% 66% 79% CD3-CD4-CD45RO+ (>31.5%) CD3+ (<40%) CD8+ (<20%) CD3+CD69+ (<15%) CD13+CD45RO- (<17.5%) 26. CD3+ (<40%) >40% 66% 75% CD3-CD45RO+ (>15%) Ratio CD13+/CD3+ (>0.675%) CD3+CD8+ (<16%) CD8+CD69- (<21%) 27. CD3+CD20-CD5- (>7.7%) >45% 61 % 80% CD4+CD13- (<20.5%) CD56-CD122- (<47%) CD4+CD45RO- (<16%) 28. CD3+ (<40%) >20% 61% 86% CD3+CD8+ (<16%) CD13+CD45RO- (<17.5%) CD3-CD20- (>58%) 29. CD13+CD45RO- (<17.5%) >70% 90% 54% CD4+CD45RA- (<16%) CD3+CD122- (42.5%) CD8+CD69- (<21 %) 30. CD3+CD8+ (<16%) >70% 84% 60% CD13+CD45RO- (<17.5%) CD4+CD45RA- (<16%) CD3+CD45RO- (<30%) CD3-CD5- (>54%) Marker combination Threshold Specificity Sensibility value 1 Blood leukocyte markers 31. CD3-CD5+ (>14.5%) >55% 66% 60% CD3-CD45RA- (>14.5%) CD3-CD44+ (>13%) CD13+ (>17.5%) CD3-CD57-CD44- (<41.3%) 32. CD3-CD45RA- (>17%) >22% 61 % 64% CD20-CD44+ (>17%) CD20-HLADR+ (>20%) CD3-CD4-CD44+ (>40.5%) CD36-HLADR+ (<5.6%) 33. CD3-CD45RA- (>14.5%) >40% 62% 64% CD3-CD45RO+ (>19%) CD20-HLADR+ (>14.5%) Blood (in italics) and endometrial leukocyte markers 34. CD57+ (>10%) >50% 76% 72% CD 14+ (>10%) CD3-CD69+ (>17.5%) CD3+ (<40%) CD4+ (<15.5%) CD3+CD8+HLADR- (<35%) 35. CD3-CD69+ (>17.5%) >33% 70% 79% CD3+ (<40%) CD4+ (<15.5%) CD3+CD8+HLADR- (<35%) Marker combination Threshold Specificity Sensibility value 1 36. CD4-CD36+ (>14.4%) >43% 70% 74% CD3-CD69+ (>17.5%) CD8+ (<20%) CD13+ (>29%) CD3+ (<40%) CD16+ (>27%) CD69+(<33%) CD5+ (<40%) 37. CD3-CD45RA- (>14.5%) >50% 73% 71% CD3-CD45RO+ (>19%) CD20-HLADR+ (>14.5%) CD14+CD44+ (>15%) CD8+ (<20%) CD5+ (<37%) CD3-CD20- (>58%) CD3-HLADR- (>54.5%) 1 Value above which a diagnosis of endometriosis is given. 2 Cutoff point established for each individual marker. TABLE 2 Examples of logistic regression models provided by endometrial or blood leukocyte markers for the identification of patients with endometriosis Marker combination B value Threshold Specificity Sensibility Number of value 1 sample tested Endometrial leukocyte markers Combination no. 1 1 CD3+ (<40%) 2 -7.9747 >.55 83% 79% 41 2. CD3-CD5- (>60%) 7.2921 3. CD13+CD45RO- -0.1410 (<17.5%) -1.6259 4. CD3-CD20- (>58%) 9.5142 Interaction of 1 to 4 Constant = 2.0516 Marker combination B value Threshold Specificity Sensibility Number of value 1 sample tested Combination no. 2 1. CD3+ (<40%) -6.7753 >.55 74% 73% 67 2. CD3-CD5- (>60%) 5.8240 3. CD13+CD45RO- -1.9298 (<17.5%) -0.0262 Interaction 1 to 4 2.8385 Constant = 2.7910 Combination no. 3 1 . CD3+CD8+ (<16%) -0.1308 >.50 84% 72% 51 2. CD13+CD45RO- -2.6688 (<17.5%) -1.1778 3. CD3+CD5+ (<37%) Constant = 3.1417 Combination no. 4 1. CD3+ (<40%) -1.6965 >.50 78% 75% 81 2. Length of menstruation -1.8160 (>7days) -1.9656 3. CD13+CD20- (<21 %) 10.3064 4. Pelvic pain 3 Constant = 3.1984 Blood leukocyte markers Combination no. 1 1. CD14+CD44+ (>15%) 0.9298 >0.55 80% 70% 140 2. CD57+ (>10%) 0.7423 3. CD3-CD45RA- (>12%) -0.8147 4. CD14+ (>10%) 0.8629 Combination no. 2 1. CD14+ (>10%) 10.5891 >.50 65% 71 % 125 2. CD57+ (>10%) 0.7326 3. CD3+CD69+ (>17.5%) 0.6899 4. CD3+HLADR+ (<4%) 1.2004 5. CD3-CD45RA- (>12%) -0.1137 Constant = -1.2062 Marker combination B value Threshold Specificity Sensibility Number of value sample tested Combination no.3 1. CD14+ (>10%) 1.1994 >.55 76% 75% 142 2. CD57+ (>10%) 0.8080 3. CD3+HLADR- (<4%) 1.3593 4. CD3-CD45RA- (>12%) -0.63 5. Pelvic pain 2.1506 6. Length of menstruation .7489 (>7d) Constant = -1.771 Combination no.4 1. CD14+ (>10%) .9727 >.50 71 % 78% 141 2. CD57+ (>10%) .4489 3. CD3+CD69+ .8129 (>17.5%) 1.3368 4. CD3+HLADR- (<4%) -0.8805 5. CD3-CD45RA- (>12%) 2.1574 6. Pelvic pain 1.5164 7. Age (>40) Constant = -1.7686 1 Value above which a diagnosis of endometriosis is given. 2 Cutoff point established for each individual marker. 3 presence of pain at any time other than menstruation and intercourse The predictive models for endometriosis were established according to the following equation: P r) = c + Bl* (markerl) + B2* (marker2) + ... Bn* (marker n) 1+ e c + Bl* (markerl) + B2.*. (marker2) + ... Bn*. (marker n) Where: P(r) = probability of having endometriosis; c = constant established for a particular combination; B = coefficient of regression; and n = total number of markers in the combination. In the present invention, a series of endometrial and peripheral blood leukocyte subpopulations for which proportions were modulated in patients with endometriosis (stage I-IV;I-II or III- IV) compared with those of normal controls, have been identified. The novelty of the present invention is to use these leukocyte subpopulations, either alone or in combination, as markers for the diagnosis of endometriosis. Moreover, risk factors for endometriosis identified amongst personal information and menstrual characteristics were shown to be of significant value when used in combination with blood or endometrial leukocyte subsets in a predictive test for endometriosis. Two methods were used for the combination of markers . Method 1 A cutoff point is established for the proportion of each leukocyte markers in order to obtain the best discrimination between patients with endometriosis and controls. The proportion obtained for each marker is compared to the cutoff point. A positive test result gives a score of 1, whereas a negative test result gives a score of 0. The diagnostic value is obtained by adding the scores of all the markers of a particular combination and converting it in percentage. The final diagnostic value is then compared to a threshold value that was established to provide the best levels of sensibility and specificity. A positive diagnosis of endometriosis is given when the final diagnostic value exceeds the threshold value established for a particular combination of markers. On the opposite, a negative diagnosis of endometriosis is given when the final diagnostic value is lower than the threshold value (see Fig. 1) . Method 2 A predictive model for endometriosis is established by including each marker of a particular combination in the following logistic regression equation: P (r) = e . c + Bl* (markerl) + B2* (marker2) + ... Bn* (marker n) -i _ c + Bl* (markerl) + B2i (marker2) + ... Bni (marker n) Where: P(r) = probability of having endometriosis; c = constant established for a particular combination; B = coefficient of regression; and n = total number of markers in the combination. The probability of having endometriosis (P(r)) is then compared to a threshold value that provides the best discriminative value. A positive diagnosis of endometriosis is given when the P(r) value exceeds the threshold value established for a particular combination of markers. Alternatively, a negative diagnosis of endometriosis is given when the P(r) value is lower than the threshold value. In the present invention, there is reported a series of 102 endometrial CD45+ leukocyte populations and 93 blood mononuclear CD45+ leukocyte populations which were shown by flow cytometric analysis to be modulated in patients with endometriosis compared with normal controls and, thus are candidate markers for the diagnosis of endometriosis (Tables 3, 4, 5, and 6) . An innovative feature of the present invention is to use these markers in combination to increase their level of sensibility and specificity in the diagnostic test . TABLE 3 Endometrial leukocyte populations proposed as good predictive markers for the identificatio of patients with endometriosis Mean proportion (% ± s.d.) of leukocyte Number of subsets samples tested Leukocyte Subsets Controls Endo Control Endo area under 3 Cutoff Specificity Sensitivity Odds (CD 4 stage I-IV ROC curve 2 point ratio CD3+ 47.7± 12.3 38.7 ±12.6 3.37x10 5 58 88 0.703 3.3X10-5 <40 84 55 6.5 (2.9-14.9) CD4+ 18.3 ±5.6 15.7 ±5.9 0.008 55 88 0.632 0.008 <17 63 63 3.0 (1.5-6.2) <15.5 72 52 2.9 (1.4-6.0) CD5+ 45.1 ±11.6 36.3 ±12.3 1.6x10" 4 466 74 0.702 0.0002 <37 80 53 4.2 (1.8-9.7) <40 73 58 3.9 (1.7-8.8) CD8+ 24.3 ±8.5 18.5 ±8.6 1.4x10 " " 54 87 0.688 0.00019 <20 74 62 4.8 (2.3-10.2) CD3+CD4+ 17.2 ±5.5 14.4 ±5.7 0.004 55 88 0.641 0.004 <15 67 57 2.7 (1.3-5.5) CD3+CD4- 29.9 ±9.6 23.6 ±9.1 1.2x10-" 55 88 0.687 0.00017 <24 80 52 4.4 (2.0-9.6) CD3-CD4- 51.1 ±14.2 60.8 ±12.6 4-lxlO 5 55 88 0.698 6.9x10-5 >61 83 50 5.1 (2.2-11.7) CD3+CD8+ 18.9 ±7.5 13.7±7.7 1.1 x 10 4 54 84 0.714 2.3x10-5 <16 70 70 5.6 (2.6-11.8) <13.5 81 54 5.3 (2.3-11.9) CD3+CD8- 26.1 ±7.8 23.1 ±7.1 0.022 54 84 0.609 <23.5 68 51 2.2 (1.1-4.7) CD3-CD8- 49.6±12.0 58.2±13.0 1.3x10-" 54 84 0.688 1.9x10-4 >53.5 70 63 4.0 (1.9-8.5) CD3+CD69+ 20.4 ±9.6 15.5 ±8.0 0.003 44 76 0.642 0.010 <15 67 53 2.4 (1.1-5.2) CD3+CD122- 41.4± 10.0 34.4±12.4 0.011 29 53 0.669 0.012 <42.5 64 76 2.9 (1.1-7.5) CD3+.ILΛDR- 38.1 ±10.3 30.6 ±12.3 2.8x10 " " 51 80 0.681 0.0005 <35 72 63 4.0 (1.8-8.5) CD3-H ADR- 46.4 ±13.0 55.6 ±13.5 1.9x10 " " 51 80 0.693 0.0002 >54.5 80 51 4.1 (1.8-9.3) Mean proportion (°Λ >±s.d.) of leukocyte Number of subsets samples tested Leukocyte Subsets Controls Endo p 1 Control Endo area under p3 Cutoff Specificity Sensitivity Odds (CI) 4 stage I-IV ROC curve 2 point ratio CD3+CD45RA+ 74±47 57±30 0018 56 85 0608 0030 <49 77 40 22 (10-47) CD3+CI.45RA- 403 ± 112 327± 120 25x10" 56 85 0684 00002 <37 69 66 47 (22-97) <35 73 60 41 (19-85) CD3-CD45RA- 314± 127 396 ±134 42x10" 56 85 0667 82x 10" >32 51 69 23 (11-46) CD3+CD45RO- 31 O± 111 250 ± 98 0002 50 73 0661 0002 <28 65 62 31 (14-66) <30 55 70 27 (13-57) CD3+CD16- 455 ± 121 372±131 21x10" 57 83 0680 00003 <38 80 49 45 (20-99) <40 75 58 42 (20-89) <475 40 80 26 (12-56) CD3+CD56- 465 ±122 385 ± 128 32x10" 56 83 0674 000053 <40 78 55 46 (21-99) Cmi CD5+ 419± II 4 333± 123 21x10" 45 74 0695 000036 <37 77 61 47 (21-109) CD3-CD5- 506±120 590± 129 0001 45 74 0690 000052 >60 82 51 46 (19-112) >54 66 66 43 (19-96) CD4+CD69- 164±48 138 ± 51 0012 37 72 0648 0012 <14 78 53 41 (16-101) CD4+CD45RA- 167±53 142±57 0010 54 85 0632 0009 <16 62 66 33 (16-67) CD8+CD69- 240 ± 79 189 ± 83 0007 30 59 0687 0004 <I8 83 53 59 (19-176) <195 76 59 51 (19-139) <21 65 68 42 (16-107) CD8+HI ADR- 233 ± 77 181 ±85 0001 49 79 0673 0001 <18 77 54 41 (18-92) CD8-HLADR- 616±95 681 ± 95 22x10" 49 79 0675 00009 >615 52 77 38 (17-83) CDΠ-CD122+ 270±98 336± 187 0031 32 58 0605 >28 64 59 25 (10-62) Mean proportion (% ± s.d.) of leukocyyttee Number of subsets samples tested Leukocyte Subsets Controls Endo pp 1 ' C Coonnttrrooll EEnnddoo aarreeaa uunnddeerr P P 33 C Cuuttooffff Specificity Sensitivity Odds (CI) 4 stage 1-1V RROOCC ccuurrvvee 22 p pooiinntt ratio CDI3-f 1)122- 471 ±144 404±150 0043 32 58 0635 0035 <46 CD20-CD5+ 446 ±120 364 ±126 0001 41 66 0681 0002 <41 <44 CD20-CDS- 520 ±128 609 ±129 0001 4 411 6 666 0 0669922 0 000000099 > >6600 77 50 36 (15-86) CD56-CD16+ 220±122 270 ± 166 0044 56 84 0571 CD56-CD16- 516 ± 122 428 ± 135 12xlθ" 5 566 8 844 0 0668877 1 188xx1100"" < <4466 71 57 33 (16-67) ratio CD3+/CD45RO+ 15 ± 10 12 ±07 0020 51 80 0626 0015 CD14+CD13- 14±09 23 ± 18 0041 21 36 0639 CD3+CD20- 444±112 369 ±135 0024 2 2 2444 4 4 4555 0 0 0666666777 0 0 0000222333 < < <444000 78 58 41 (14-123) CD3-CD20- 523 ± 115 610 ±149 0016 22 2444 44 4555 00 0666666999 00 0000222222 >> >555888 83 53 62 (18-212) >>5566 70 60 45 (14-135) CD3-CD4-CD45RA+ 40 I ± 137 349 ± 151 0046 51 79 0618 0023 CD3-CD4-CD45RA- 579 ±141 633 ±152 0042 51 79 0620 0021 CD3+CD8+CD69- 404±102 351 ± 117 0039 2 299 5 566 0 0663355 0 0004422 < <334455 75 52 34 (I 2-92) CD3+CD8+HLADR- 395 ±92 336± 115 0003 4 488 7 744 0 0666655 0 0000022 < <3355 72 55 35 (16-77) CD3+CD8-HLADR- 431 ± 75 463 ± 109 4 4 4888 7 7 7444 0 0 0666000333 > > >444777 6 699 5 500 2 222 ( (1100--4477)) CD3-CD8-HLADR- 767±89 803 ± 79 0021 4 4 4888 7 7 7444 0 0 0555999777 CD14+CD13-CD16b- 198 ±164 334 ± 229 0026 1199 3366 00770033 00001144 >>2233 68 69 49 (14-163) >145 53 83 46 (14-156) Mean proportion (% ±s.d.) of leukocyte Number of subsets samples tested Leukocyte Subsets Controls Endo p 1 Control Endo area under 3 Cutoff Specificity Sensitivity Odds (CI) 4 stage I-IV ROC curve 2 point ratio CD4H D14- 207 ± 78 148 ± 58 0014 14 23 0738 0017 <166 79 65 69 (15-320) CD4-f D14- 757 ± 81 810±57 0025 14 23 0711 0033 >76 57 83 63 (14-287) CD4+IILADR- 160±57 123 ±41 0018 14 30 0715 0023 <17 57 87 87 (19-386) CD13-CD69+ 542 ±143 423 ± 182 0039 14 30 0705 0030 <51 71 70 58 (14-236) CD13+CD45RO- 221 ± 88 155 ± 107 0009 25 54 0746 46x 10" <175 76 70 75 (25-223) <21 52 80 42 (15-118) CD56-CD122- 490 ±122 428 ±158 29 51 0631 <47 65 69 36 (14-93) CD3+CD69- 264 ± 70 233 ± 84 0036 44 76 0612 0041 <27 57 67 22 (10-48) CD4+CD45RO- 149±53 130±62 47 74 0615 0034 <16 51 72 26 (12-56) CD56 1)122+ 35±22 26±17 29 51 0651 0025 <30 55 73 33 (13-85) CD3-CD56+CD16+ 82±38 68±40 0049 53 78 0615 0026 <65 72 55 30 (14-62) CD3-CD56+CD122+ 36±20 26± 18 0033 27 49 0638 0048 <27 63 65 32 (12-85) CD14+CD13+ 38±19 30±26 21 36 0672 0032 <23 81 53 48 (13-169) C!)3+CI)20-CT)5+ 930 ± 27 880± 138 24 40 0659 0034 <915 79 55 46 (14-149) CD4-CD13+CD16+ 31 O± 140 224 ±189 16 36 0699 0023 CD69+ 418±129 388± 178 43 78 0557 <33 81 41 30 (12-74) ratio CD H+/CD3+ 056 ± 054 078 ±071 46 78 0596 >068 80 40 26 (11-61) Mean proportion (% ± s.d.) of leukocyte Number of subsets samples tested Leukocyte Subsets Controls Endo j f Control Endo area under Cutoff Specificity Sensitivity Odds (CI) 4 stage 1-1V ROC curve 2 point ratio CD3-CD20-CD5- ;0±69 907±52 24 40 0598 >84 37 90 54 (14-203) CD3+CD20-CD5- 51 ±23 95±136 24 40 0683 0015 >77 87 50 99 (21-481) CD4+C 1 ) 13- 175 ±69 158 ± 54 36 63 0594 <205 42 86 43 (16-113) CD3+CD44- 417± 120 383 ±136 31 56 0596 <378 74 50 29 (11-75) CD56+ 262±125 302±172 57 87 0562 >32 41 29 (13-65) CD13-CD45RO+ 214 ± 87 263±113 25 54 0625 >28 80 45 32 (10-98) CD56+CD69- 196± 129 243±143 33 53 0610 >26 85 40 34 (11-102) CD13-CD16+ ;o±75 67±33 39 71 0562 <6 72 51 26 (11-61) CD56+ D122- 218±1I8 287 ± 173 29 51 0621 >19 59 71 31 (12-79) >18 55 72 33 (13-85) CD3+CD4-CD69+ 372±100 340 -± 138 34 66 0572 <335 73 49 26 (11-64) CD4-CDΠ-CD16+ 15±36 87± 101 16 36 0642 <71 75 58 42 (11-156) CD4-CD13-CD16- 545 ± 130 622±214 16 36 0655 >65 81 56 54 (13-223) Mean proportion (% ± s.d.) of leukocyte Number of subsets samples tested Leukocyte Subsets Controls Endo p' Control Endo area under Cutoff Specificity Sensitivity Odds (CI) 4 stage I-IV ROC curve 2 point ratio CD14+CD13+CD16b+ 1 1.0 ± 12.5 6.5 ± 6.2 19 36 0.616 <16 32 94 7.8 (1.4-43.9) CD4-CD69- 40.7 ± 12.9 46.4 ± 18.6 37 72 0.593 >47 76 47 2.8 (1.2-6.7) CD3-CD45RO+ 23.0 ± 12.5 27.1 ± 16.3 50 73 0.556 >15 34 80 CD4-CD69+ 39.3 ± 1 1.8 37.7 ± 18.0 37 72 0.462 <35 68 46 CD3-CD4-CD45RO+ 41.3 ± 18.1 43.4 ± 20.9 43 72 0.530 >31.5 33 71 >56 81 28 P value (when < 0.05) obtained in a student "t" test when mean proportion found in patients with endometriosis stage I-IV was compared to normal controls. Discriminative value of each marker established by area under ROC curve. P value (when < 0.05), significance of area under ROC curve. Confidence interval for odds ratio. rei -t-pnt-j -C-J. U1UUU ιeui-υu ) ' e u u.-La.i-. t. JJLJ.S j jiu uscu c-» yuuu ieuiu iv e iiiciJ. i c i. a -.<- )±. -.lie: ident ification of patients with endometriosis Mean proportion (% ± s d.)of Number of samples leukocyte subsets tested Leukocyte Subsets Control Endo. P 1 Control Endo. area under P 3 Cutoff Specificity Sensitivity Odds (CI) 4 Stage I-IV Stage MV curve 2 point ratio CD3+ 666 ± 85 645 ± 87 0032 132 172 0570 0037 CD8+ 173 ± 52 164±48 129 172 0549 <189 33 77 17 (10-28) CD13+ I60±60 176±65 0039 122 155 0575 0032 >175 63 51 18 (11-29) CDI4+ II 8±49 I34±60 0020 124 167 0575 0029 >10 45 71 20 (13-33) CD20+ 57±31 48±23 0006 124 162 0582 0017 <6 39 74 18 (11-30) CD36+ 157±68 172±73 112 140 0560 >19 77 37 21 (12-36) CD44+ 171 ±56 191 ±65 0009 113 148 0585 0018 >185 61 51 17 (10-27) CDS7+ 80±39 92±49 0023 114 148 0569 >10 75 39 18 (11-31) CD69+ 194±82 210±71 109 144 0590 0014 >215 71 45 20 (12-34) CD122+ 292±84 312±117 122 166 0567 >34 74 42 21 (12-34) CD3+CD5+ 666±85 637±104 0017 115 146 0586 0017 <69 44 70 18 (11-29) CD3+CD45RA- 393 ±95 372±83 0044 124 168 0583 0015 <42 40 72 17 (11-28) CD3+CD56- 653 ± 88 632 ± 86 0035 126 169 0571 0037 <68 42 71 18 (11-29) CD3+CD57- 637±83 607 ± 97 0009 113 146 0592 0011 <67 40 77 23 (13-39) CD3+CD69- 601 ± 96 579±92 107 141 0584 0023 CD3+CD122- 622 ± 84 596±98 0021 121 164 0578 0024 <58 69 42 16 (10-27) CD3+HLADR+ 39±14 35±12 0006 121 154 0601 0004 <4 40 77 22 (13-37) CD3-CDS+ 155±53 183 ± 84 0003 115 146 0619 0001 >145 50 72 25 (15-42) CD3-CD16+ 233 ± 79 254 ± 87 0036 123 166 0572 0037 CD3-CD44+ 131±53 I56±63 0001 111 143 0615 0002 >115 44 73 22 (14-37) CD3-CD57+ 42±25 50±31 0023 113 146 0573 0044 CD3-CD69+ 142±56 164±65 0006 107 141 0602 0006 >175 75 41 21 (12-36) CD3-CD45RO+ 160±56 18 I±68 0006 117 148 0595 0008 >19 77 44 26 (15-45) CD3-CD4- 311 ±95 333 ±90 0043 132 171 0566 0050 CD3-CD8- 334±92 355± 100 122 166 0568 0050 CD3-CD45RA- 142±52 162±62 0004 124 168 0595 0006 >145 61 60 23 (14-37) CD3-CD56- 216±63 231 ± 70 126 169 0562 >25 73 39 17 (10-28) CD4-CD13+ 145±60 163±68 0030 108 131 0586 0023 >165 69 51 24 (14-41) CD4-CD36+ 138±74 190±62 0037 13 21 0771 0009 >19 92 62 195 (21-1799) Mean proportion (% ± s .d.) of Number of samples leukocyte subsets tested Leukocyte Subsets Control Endo. p' Control Endo. area under P 3 Cutoff Specificity Sensitivity Odds (CI) 4 Stage I-IV Stage I-IV curve 2 point ratio CD4-CD69+ 160±61 181 ±65 0021 94 120 0603 0009 >19 75 45 24 (13-43) CD4-CD45RO+ 234 ± 67 270 ± 77 0043 27 50 0620 CD4-CD45RA- 227 ± 65 244 ± 83 125 168 0562 CD8-C D44+ 168 ± 55 189 ± 67 0017 88 119 0588 0030 CD8-CD44- 664 ± 70 644 ± 66 0039 88 119 0584 0040 <68 48 70 21 (12-37) CD13+CD44+ 133 ± 53 154±59 0006 96 121 0605 0008 >11 38 76 19 (11-34) CD13+HLADR+ 130±53 147±6 I 0024 108 135 0581 0031 >155 71 42 18 (11-31) CD13+CD16- 14±09 37 ± 46 0005 21 37 0727 0004 >2 76 57 42 (13-139) CD13-MI ADR+ 82±32 70±30 0005 108 135 0619 0001 <8 50 70 24 (14-40) CD13-CD44- 799 ± 62 769 ± 86 0004 96 121 0606 0007 <82 39 76 20 (11-36) CD14+HLADR+ 110±46 127±59 0009 109 147 0587 0017 >95 48 70 22 (13-36) CD14+CD44+ 107±49 129±57 0003 85 123 0612 0006 >15 85 33 27 (13-54) CD14+CD45RO+ 122±44 143±61 0004 102 118 0586 0028 CD14+CD16- 07±08 27±54 0023 23 41 0603 CD14+CD122- 04±03 24 ±52 0029 14 34 0648 >07 86 47 53 (10-275) CD14-III ΛDR+ 103 ±33 91 ±27 0001 109 147 0611 0002 <85 69 47 20 (12-33) CD14-CD44- 835 ± 53 816±63 0023 85 123 0588 0032 <80 79 37 22 (12-42) CD20+HLADR+ 52±29 43±2I 0005 106 141 0596 0010 <55 39 75 19 (11-33) CD20+CD44- 50±28 42±22 0033 95 126 0571 <4 62 55 20 (12-34) CD20-CD44+ 144±51 173±62 00004 95 126 0636 0001 >17 74 47 25 (14-44) CD20-CD69+ 164±43 210±62 0016 14 29 0719 0021 >16 57 76 42 (11-163) CD20-HLADR+ 154±49 176±69 0004 106 141 0591 0014 >145 49 70 22 (13-37) CD20-CD44- 795 ± 51 775 ± 61 0008 95 126 0590 0022 CD20-CD69- 794 ± 67 753 ± 65 0065 14 29 0711 0027 <756 71 52 116 (23-570) CD36-HLADR+ 78±32 64±20 00005 95 121 0629 0001 <56 77 40 22 (12-40) CD56-CD69+ 181 ±42 218±67 0083 13 23 0667 >205 85 52 60 (11-333) CD56-CD122+ 195 ± 57 214±86 0028 113 154 0578 0030 >23 74 40 20 (12-33) CD56-CD69- 679±53 627 ± 94 0075 13 23 0671 0093 CD56-CD122- 677±83 647 ± 115 0014 113 154 0586 0017 <645 67 51 21 (13-35) CD57-CD44- 743 ± 69 717±77 0015 84 117 0602 0014 <76 44 71 19 (11-35) CD3+CD57-HLADR+ 43 ± 15 40±14 96 118 0584 0034 <37 72 43 19 (11-35) CD3-CD4-CD44+ 344±110 381 ±126 0032 88 112 0577 >405 69 45 18 (10-33) CD3-CD56+CD16- 12±06 14±10 0023 121 163 0553 CD3-CD56-CD122- 234 ± 111 212 ± 114 113 148 0581 0024 CD3-CD57-CD44- 487± 101 445 ±103 0004 84 114 0610 0008 <413 80 41 28 (I 4-53) Mean proportion (% ± s..d.) of Number of samples leukocyte subsets tested Leukocyte Subsets Control Endo. P 1 Control Endo. area under P 3 Cutoff Specificity Sensitivity Odds 4 (CI) Stage I-IV Stage l-IV curve 2 point ratio CDI4+CD20+CD44- 0.2 ± 0.2 0.1 ± 0.1 0.036 65 75 0.613 0.037 CD14+CD20-CD44+ 91.8 ± 4.6 93.5 ± 3.5 0.016 65 75 0.614 0.021 >95 75 43 2.3 (1.1 -4.7) CD14+CD20-CD44- 5.9 ± 4.5 4.6 ± 3.3 0.037 65 75 0.604 0.035 <3 75 43 2.3 (1.1-4.7) CD14-CD13-IILADR+ 8.7 ± 3.5 7.6 ± 2.6 0.021 77 82 0.600 0.029 <7 71 45 2.1 (1.1 -4.0) Ratio CD 13+/CD3+ 0.25 ± 0.12 0.29 ± 0.14 0.025 121 153 0.583 0.018 >0.30 71 43 1.8 (1.1-3.0) Ratio CD13+CD8+ 1.04 ± 0.55 1.20 ± 0.69 0.039 1 14 153 0.574 0.039 Ratio CD 14+/CD3+ 0.19 ± 0.10 0.22 ± 0.12 0.01 1 123 165 0.575 0.028 >0.14 40 74 1.9 (1.1-3.1 ) Ratio CD14+/CD8+ 0.78 ± 0.47 0.91 ± 0.55 0.040 1 16 164 0.574 0.036 1 P value (when < 0.05) obtained in a student "t" test when mean proportion of leukocyte subsets was compared between patients with endometriosis (stage I-IV) and normal controls. * Discriminative value of each marker established by area under ROC curve. 3 P value (when < 0.05), significance of area under ROC curve. 4 Confidence interval for odds ratio. TABLE 5 Endometrial leukocyte populations used as markers to discriminate between patients with endometriosis stage I-II or stage III-IV and normal subjects Mean proportion (% ± s.d.) of Number of samples leukocyte subsets tested _ , Endo „ . . Endo „/ . .,. .ndo Endo area under Cutoff SpecificSensitivOdds Leukocyte Subsets Controls _ . ¥ ¥¥ p stage Ill- p 2 Control P (CI) 4 stage I-II I-II III-IV ROC curve point IV ity ity ratio CD3-CD44- 500 ±129 505 ± 119 618 ±166 0015 31 43 13 0766 0007 >48 52 92 129 (14-1138) CD3+HLADR+ 83 ±57 61 ± 26 0013 76±33 51 57 23 0587 <55 67 483 CD3+CD45RO+ 149±84 120±56 0043 132 ±95 50 50 23 0565 >15 373 71 CD4+CD13+ 23 ± 19 22± 17 14 ± 11 0025 36 44 19 0658 <17 50 79 38 (10-138) CD13+HLADR+ 51 ±21 53±23 36 ± 13 0017 27 37 12 0740 0021 <5 54 92 13 (14-1172) K wm CD14-HLADR+ 147±75 102±39 0049 108 ± 51 14 24 4 0679 <13 57 75 CD56+HLADR+ 28±17 15 ± 07 0011 46±33 16 17 5 0741 0018 <17 75 71 72 (15-336) CD56-CD44- 635 ±130 635 ±134 511 ±204 0023 30 41 12 0708 0041 <65 58 83 68 (12-375) CD3-CD4+CD45RΛ- 15 ±09 15 ± 09 10±07 0022 51 54 25 0668 0021 <07 84 40 34 (I 1-107) CD3+CD8-HLADR+ 159 ± 81 158± 86 236 ±121 0011 48 52 22 0701 0009 >148 55 73 33 (11-101) CD3-CD8+HLADR- 95±65 86 ± 61 61 ± 52 0036 48 52 22 0691 0013 <5 78 59 50 (16-154) CD3+CD56+CD16+ 30±34 29±32 60 ± 57 0027 53 56 22 0670 0024 >18 49 77 35 (10-103) CD3+CD56-CD16+ 66±75 70±62 II 0 ± 91 0350 53 56 22 0698 0009 >9 84 50 54 (17-173) CD3+CD56-CD16- 870 ± 81 872 ± 71 795 ±129 0016 53 56 22 0695 0010 >83 81 50 41 (14-125) Mean proportion (% ± s.d.) of Number of samples leukocyte subsets tested _ . Endo -. _ . i Lndo / . ... _ . . Endo Endo area under Cutoff SpecifiSensitivOdds Leukocyte Subsets Controls . , „ p stage Ill- trol .., .... y P stage I-II r = v P 2 Con ROC curve point city ity ratio (CI) 4 CD3+CD56-CD44+ 70±61 61 ±48 132± 105 0025 28 38 12 0721 0034 <5 42 92 CD3+CD56-CD44- 871 ±61 885 ±62 780 ±139 0049 28 38 12 0668 >81 83 58 70 (15-337) CD3-CD56-HLADR- 297± 156 427 ± 196 0050 293 ±213 15 17 6 0706 0047 <36 73 59 CD3+CD56-CD122+ 121 ±75 119±63 217± 140 0005 27 33 16 0715 0024 >20 87 50 67 (14-317) CD3+CD56-CD122- 819±82 830±68 680±217 0024 27 33 16 0715 0024 >765 71 63 40 (11-155) H H CD4-CD16- 515 ± 129 591 ±127 0033 493 ±214 24 31 11 0659 0045 <58 76 47 m H CΛ CD14+CD13+CD16b- 627 ± 207 562 ±258 382 ±264 0011 19 26 10 0753 0035 >61 69 80 88 (13-574) a w w CD16+ 274 ±124 300±162 354±195 58 62 23 0603 >39 85 39 35 (12-105) H >27 50 51 CD45RA+ 283 ±91 276 ± 90 279 ± 160 56 60 25 0562 <235 75 48 28 (10-75) t- 1 to CD45RO+ 384± 136 38 O± 155 446 ± 181 50 54 24 0620 >52 88 43 44 (13-144) O CD13+ 248±127 255±147 313±203 47 57 22 >29 68 34 1 P value obtained in a student "t" test when mean % of leukocyte subsets found in patients with endometriosis stage I-II was compared to normal controls 2 P value obtained m a student "t" test when mean % of leukocyte subsets found m patients with endometriosis stage III-IV was compared to normal controls. TABLE 6 Peripheral blood leukocyte populations used as markers to discriminate between patients with endometriosis stage I-II or stage III-IV and normal subjects Mean proportion (% ± s.d.) of Number of samples leukocyte subsets tested F ri Endo Endo Endo area under Cutoff Specific- Sennssiittiivv- Odds Leukocyte Subsets Controls . . „ p stage Ill- p 2 Control (CI)" stage I-II r B iy I-II III-IV ROC curve point ity ity ratio CD3-CD57+CD44- 125 ±62 149 ±75 0030 122± 69 84 81 33 0586 >156 74 43 21 (11-41) CΛ G CD14-CD13-HLADR- 856±50 876±39 0016 859 ± 33 77 54 28 0637 0008 >883 74 47 26 (11-53) W CΛ H CD14-CD20+CD44- 57±29 47±26 0043 56±27 65 50 23 0607 0049 <45 66 53 22 (10-47) H G HLADR+ 216±60 217±66 238 ± 67 0037 120 102 52 0586 >245 73 45 22 (11-45) H W CD3+CD8- 499±85 490±92 463 ± 88 0012 122 113 53 0654 0002 <515 48 85 52 (22-121) CD3+CD44- 602±86 592±88 572 ± 80 0047 111 98 45 0605 0047 <612 51 73 28 (13-61) CD3+HLADR- 633 ± 87 630 ± 91 596 ± 79 0009 121 102 52 0628 0010 <635 55 73 32 (15-66) CD3-HLADR+ 181 ±58 187±65 208±69 0010 121 102 52 0607 0030 >215 73 45 23 (11-46) OS CD3+CD16+ 93 ± 133 84± 121 60±43 0016 123 113 53 0528 >37 31 81 CD3-CD57- 283±69 290±72 323 ± 95 0004 113 101 45 0644 0007 >28 53 71 27 (12-57) CD4-HLADR+ 198±60 198±62 220 ± 66 0036 112 96 49 0588 >193 48 67 CD4+CD45RA- 314± 81 297±72 277 ± 85 0006 125 114 54 0625 0010 <295 60 60 23 (12-45) CD4+CD45RO+ 195±56 205±64 162±43 0029 27 31 19 0645 <198 50 90 85 (16-445) CD13-CD16+ 189± 82 168± 87 143 ±31 0035 21 28 9 0698 <13 86 56 75 (12-451) Mean proportion (% ± s.d.) of Number of samples leukocyte subsets tested „ . Endo Endo ; . ... Endo Endo area under Cutoff Specific- Sen issiittiivv-- Odds Leukocyte Subsets Controls p stage Ill- p 2 Control stage I-II I-II III-IV ROC curve point ity (CI) 4 IV ity ratio CD14-CD69- 810 ± 61 774 ± 91 760±67 0031 20 32 14 0729 0025 <82 50 86 60 (11-340) CD20-HLADR- 793±52 785±70 771 ± 66 0025 106 97 44 0573 <803 41 72 a CD57-CD44+ 172±55 183 ±64 197±77 0046 84 82 35 0573 >216 80 40 27 (11-67) a CD3-CD4+CD45RA- 69±44 64±40 54 ± 35 0043 122 109 50 0587 <55 59 61 23 (11-45) H H CD3-CD8-CD44+ 372± 119 372 ±127 432±126 0019 87 80 32 0588 >405 54 63 H W CD3-CD57-CD44+ 385 ± 121 403 ± 125 440 ±127 0029 84 81 33 0575 >44 CΛ 63 58 ffl m CD14+CD20+CD44+ 21 ± 13 21 ± 15 14±07 0001 65 52 23 0647 <21 39 91 64 (13-309) H r g tiP value obtained in a student "t" test when mean proportion found m patients with endometriosis stage I-II was compared to normal controls ro : P value obtained in a student t test when the % leukocyte subsets found in patients with endometriosis stage III-IV was compared to normal controls σ. Cutoff points established for each individual marker are presented in Table 3, 4, 5, 6 and threshold value established for a particular marker combination are presented in Table 1. Any other cutoff points or threshold values providing a valuable diagnostic test for endometriosis are meant to be included in the present invention In accordance with a preferred embodiment of the present invention, there is provided a series of 34 different combinations of endometrial leukocyte markers (Tables 1 and 2) , 7 combinations of blood leukocyte markers (Tables 1 and 2) and 4 combinations of endometrial and blood leukocyte markers providing a diagnostic test with levels of sensibility and specificity up to 89 and 90%, respectively. The different marker combinations of the present invention may serve several important clinical needs. Hence in the general population, these markers could be used to evaluate the risk factor to develop endometriosis or to identify women with high likelihood of suffering from the disease. Furthermore in patients with endometriosis, these markers could serve to monitor the disease or to give a prognosis. Study subjects and samples Uterine endometrial tissues were obtained from 146 subjects undergoing laparoscopy or laparotomy. The experimental group was formed of up to 88 subjects with endometriosis stage I-IV confirmed by laparoscopy or laparotomy and the control group consists of up to 58 healthy subjects who underwent surgery for tubal ligation (or reanastomosis) and had no clinical evidence, nor family history of endometriosis. Table 7 gives details concerning the age, menstrual cycle and indication of laparoscopy or laparotomy for the subjects included in experimental and control groups. TABLE 7 Descript ion of the experimental groups used in the analysis of endometrial leukocyte populations Percentage of patients* Menstrual Indication of laparoscopy cycle Experimental Number Mean age 1* 2* ES LS ligation HysterectoDiagnosOther** groups of ± s.d. or my and/or tic subjects reanasovariectomy laparostomosis copy Controls 58 34.2 ± 5.3 54.5 % 45.5% 100% Endometriosis Stage I-IV 88 34.4 ± 6.8 47.0% 53.0% 21.6% 22.7% 52.3% 3.4% Stage I-II 63 34.4 ± 7.3 50.9% 49.1% 28.6% 22.2% 47.7% 1.5% Stage III-IV 25 34.4 ± 5.4 36.4% 63.6% 4.0% 24.0% 64.0% 8.0% 1 Early secretory (days 14-21) 2 Late secretory (days 22-28) * % patients among control or endometriosis groups Endometrial biopsies were taken with a Pipet 10 Curette (Milex) (approximately 0.5g of tissue). All samples were harvested in the secretory phase (day 14- 28) of the menstrual cycle as confirmed by histological evaluation. The samples were collected into sterile RPMI-1640 medium (Gibco) supplemented 15 with 2% heat-inactivated fetal calf serum (Bio-Media) and 1% penicillin-streptomycin and kept at 4°C until cell isolation. Blood samples were obtained from up to 172 subjects with endometriosis (stage I-IV) confirmed by 20 laparascopy or laparotomy and from up to 132 healthy subjects with no evidence of endometriosis at surgery, and no family history of endometriosis. Blood samples (30 ml) were collected in heparin-tubes (Vacutainer™, Becton Dickinson) and kept at 20°C until mononuclear 25 cell separation. The age, menstrual dating and indication for laparoscopy of the subjects included in the study are given in Table 8. TABLE 8 Description of the subjects included in the study of peripheral blood mononuclear leukocytes Percentage of patients* Menstrual cycle Indication of laparoscopy Experimental Number Mean age Prolife- Secre- , . * * ligation Hysterecto- Diagnos- Other groups of ± s.α. rative tory or my and/or tic subjects reanas- ovariectomy laparos- tomosis copy Control 132 34.30 ± 43.8 % 56.2% 100% 5.5 Endometriosis Stage I-IV 172 36.40* 42.8% 57.2% 22.1% 33.7% 38.9% 5.3% Stage MI 116 35.96 ± 41.0% 59.0% 31.1% 30.2% 37.1% 1.6% 6.39 Stage III-IV 56 34.30 ± 46.2% 53.8% 3.6% 41.1% 42.9% 12.4% 5.5 * % of patients amongst control or endometriosis groups Stromal cell preparation from endometrial samples Endometrial tissue samples were mechanically disrupted with a Pyrex™ glass Broeck tissue grinder 10 (Fisher) to obtain a single cell suspension. Stromal cell fraction was isolated by filtration through a 250 μm stainless steel sieve (Millipore) to retain the glandular fraction and was washed twice with 10 ml phosphate buffered saline (PBS) (Sigma) containing 1% 15 BSA (Boehringer Mannheim), 0.1% sodium azide (Fisher) (thereafter called PBS washing buffer) . Isolation of mononuclear cells from peripheral blood Blood samples were diluted 1:1 with Hank's Balanced Salt Solution (HBSS) (Gibco) , layered on an 20 equal volume of Ficoll-Paque™ (Pharmacia Biotech) and centrifuged at 1500 rpm for 40 minutes at room temperature. Leukocytes were isolated at the interface of Ficoll and HBSS and they were washed in 50 ml of HBSS. Contaminating red blood cells were 25 lysed with 6 ml of ammonium chloride solution (0.15M) (6 minutes at room temperature) . The peripheral blood mononuclear cells were then washed twice in 10 ml PBS and resuspended in PBS washing buffer. Endometrial and peripheral blood leukocyte surface antigen staining Endometrial stromal cells or peripheral blood mononuclear cells were distributed in 5 ml tubes (1 to 1.5 x 10 6 cells/tube) or in 96 well plates (5 x 10 5 cells/well) , respectively and incubated in the presence of 0.1 μg of human γ-globulin for 5 minutes at room temperature. The cells were then incubated 30 minutes in the dark (at room temperature for endometrial cells and at 4°C for peripheral blood mononuclear cells) with a panel of 4 different mouse monoclonal antibodies (MAbs) in a total volume of 100 μl . The cell samples were stained with mouse anti- human CD45 MAbs conjugated to peridinin chlorophyl protein (PerCP) and with several sets of three different mouse MAbs labeled with distinct fluorochromes (fluorescein isothiocyanate -FITC-, phycoerythrin -PE or with phycoerythrin-texas red - ECD-) directed toward cell surface markers for specific cell populations such as T lymphocytes, B lymphocytes, NK cells, macrophages and/or activation markers (Table 9) . TABLE 9 Description of mouse monoclonal antibodies used for immunophenotyping Specificity Clone Isotype Supplier Fluoro- chroπie CD3 HIT3A mouse IgG2a Beckman/Coulter ECD CD4 SK3 mouse IgG 1 Becton Dickinson PE CD5 BL1A mouse IgG2a Beckman/Coulter FITC CD8 SKI mouse IgGl Becton Dickinson PE CD13 SJ1D1 mouse IgG 1 Beckman/Coulter RPE CD14 RM052 mouse IgG2a Beckman/Coulter PE CD16 NKP15 mouse IgGl Becton Dickinson FITC CD16B 1D3 mouse lgM Beckman/Coulter FITC CD20 H299 mouse IgG2a Beckman/Coulter RD1 CD36 SMf mouse IgM Sigma RPE CD44 L178 mouse IgG 1 Becton Dickinson FITC CD45 2D1 mouse IgGl Becton Dickinson PerCP CD45-RA ALB 1 1 mouse IgGl Beckman/Coulter FITC CD45-RO UCHL1 mouse IgG2a Beckman/Coulter FITC CD56 N901 (NKH-1) mouse IgGl Beckman/Coulter PE CD57 VC1 1 mouse IgM Sigma RPE CD69 L78 mouse IgGl Becton Dickinson FITC CD 122 2RB mouse IgGl Beckman/Coulter FITC HLA-DR L243 mouse IgG2a Becton Dickinson FITC Table 10 below lists the distribution of the antigens listed in Table 9. TABLE 10 Main distribution of antigens Antigen Mam Cell Distribution CD3 Expressed on all mature T cells associated with TCR complex ( α/b , γ/δ ) CD4 Expressed on T helper lymphocytes It can be also expressed on cells of the monocyte/macrophage lineage CD5 Found on all mature T lymphocytes and a subset of B lymphocytes CD8 Found on a subset of T lymphocytes called suppressor/cytotoxic T cells Antigen Main Cell Distribution CD13 Detected on most cells of myeloid origin polymorphonuclear cells or cells of the monocyte/macrophage lineage. Member of metalloproteinase family CD14 Expressed strongly on the surface of monocytes Found on most tissue macrophages Weakly expressed on the surface of granulocytes and B lymphocytes Receptor for lipopolysacharride (LPS) and LPS binding protein CD16 Expressed mainly on NK cells, monocytes macrophages and polymorphonuclear leukocytes Low affinity receptor for IgG CD 16b Found on granulocytes including polymorphonuclear cells (PMN) CD20 Present on all B lymphocytes CD36 Expressed on platelets, monocytes or macrophages, microvascular endothelial cells, mammary endothelial cells, during stages of erythroid cell development CD44 Widely expressed on the surface of most cell types. Including most leukocytes and epithelial cells. Family of core/link peptidoglycan CD45 Present on the surface of all leukocytes CD45RA Isoforms of CD45 Found on naive/resting T cells Also expressed on B lymphocytes and monocytes CD45RO Isoforms of CD45 expressed on memory/activated T cells also expressed on monocytes CD56 Marker for NK cells Can also be found on a population of T lymphocytes CD57 Found on a subset of cells with natural killer activity CD69 Expressed on activated leukocytes including T cells, B cells, NK cells, neutrophils, eosinophils and cells of the monocyte/macrophage lineage. Activation marker detected early after cell activation CD 122 Expressed on NK cells B, T lymphocytes or monocytes/macrophages Component of the IL-15 receptor HLADR+ HLA class II molecule Found on antigen presenting cells or on other cells upon activation such as T cells. Blood cells were washed twice with 0.15 ml of PBS washing buffer. Endometrial cell samples were incubated with a red blood cell lysing solution, (FACS™ Lysing Solution, Becton Dickinson) for 10 minutes at room temperature in the dark and washed with 3 ml of PBS washing buffer. Endometrial and blood cells were fixed in 1% paraformaldehyde (diluted in PBS) at a concentration of 1 x 10 6 cells/ml and kept at 4°C in the dark until the immunofluorescence reactivity was determined by flow cytometry. Flow cytometry analysis The immunofluorescence reactivity was carried out on a Coulter EPICS XL™ flow cytometer (Coulter Corporation, Hialeah, FL) equipped with an argon laser operating at 488 nm, 15 mW and detectors at 525, 575, 610, and 675 nm. Calibration of the flow cytometer parameters for forward scatter, side scatter and fluorescence were the same for all the samples. Cells expressing CD45 pan leukocyte antigen were gated using the Coulter system II software. The percentage of cells bearing markers for T, B lymphocytes, macrophages or NK cells and/or activation markers was evaluated within the CD45 positive populations only. A minimum of 6000 CD45+ cells were analyzed for each sample . Use of leukocyte markers in a diagnostic test for endometriosis A cutoff point was established for the proportion of the endometrial or blood leukocyte subpopulations identified as diagnostic markers. The value obtained for each marker is compared to the cutoff point (Fig. 1) . A positive result was given when the proportion of a particular leukocyte subset fulfills the condition established by the cutoff point (for example < 40% for CD3+ cells) . When these markers are used in combination, a positive result for each marker gives a score of 1, whereas a negative result gives a score of 0. A diagnosis of endometriosis is given, when the final diagnostic score obtained from adding the results of all the markers of a particular combination is higher than a predetermined threshold value. The levels of sensibility and/or specificity measured for the marker combination represents the number of positive test results obtained among the patients already confirmed with endometriosis and the number of negative test results among the subjects within the control group, respectively. Endometrial and blood leukocyte markers can be used in combinations in logistic regression model : P ( r) = e_ c + Bl* (markerl) + B2* (marker2 ) + . . . Bn* (marker n) -ι , _ c + Bl* (markerl) + B2.*_ (marker2 ) + . . . Bn*_ (marker n) Where : P(r) = probability of having endometriosis c = constant established for a particular combination B = coefficient of regression n = total number of markers in the combination The probability of having endometriosis (P(r)) is then compared to a threshold value that provides the best discriminative value. A positive diagnosis of endometriosis is given when the P(r) value exceeds the threshold value established for a particular combination of markers. Alternatively, a negative diagnosis of endometriosis is given when the P(r) value is lower than the threshold value. Results Endometrial and blood leukocyte subsets defined as good potential markers for the diagnosis of endometriosis are presented in Tables 3 and 4 respectively. Selection of these markers was done on the basis of 1) a significant difference in the mean proportion of leukocyte subsets between patients with endometriosis (stage I-IV) and control groups; 2) several endometrial and blood leukocyte markers were also selected according to the area under the ROC curve (>0.500), an indication of the discriminative value of the markers. The ROC curve allowed the determination of one or more cutoffs (proportion % of leukocyte subpopulations) that best discriminate between patients with endometriosis (stage I-IV) and normal controls. In an attempt to use these differences for identifying patients with endometriosis, a positive test result was given when the proportion measured for a particular leukocyte subset fulfills the condition established by the cutoff point (for example < 40% for CD3+ cells) . The levels of specificity and sensibility were calculated for each marker used alone to diagnose endometriosis and are presented in Table 3 (for endometrial leukocyte markers) and Table 4 (for blood leukocyte markers) . Moreover, 3) a significant odds ratio calculated with a particular cutoff point gave an additional indication that the leukocyte markers selected in Tables 3 and 4 are associated with an increased risk to develop endometriosis and can, thus, be used for identifying women with high likelihood of suffering from endometriosis. The mean proportion of some endometrial (Table 5) and blood (Table 6) leukocyte subsets was found to be significantly modulated only in patients with stage I-II endometriosis or with stage III-IV endometriosis when compared to normal controls. These markers remain good candidates for a diagnostic test for endometriosis, but their use may be limited to a specific stage of the disease. Several of endometrial and blood leukocyte markers were found to be even more reliable as diagnostic markers when they are analyzed in combination with other markers. Table 1 gives a series of 33 combinations in which endometrial or blood leukocyte markers are used in a diagnostic test for endometriosis. For each marker, a positive test result (as described above; see also Fig. 1) gives a score of 1, whereas a negative test result gives a score of 0. The final diagnostic value obtained from adding the scores of all the markers of a particular combination is then compared to a threshold value, which is indicated in Table 1. A diagnosis of endometriosis is given, when the diagnostic value exceeds the threshold value established for each set of combination markers. The use of leukocyte marker subsets in combination in this new method clearly improves the levels of sensibility and/or specificity for diagnosing endometriosis. Table 1 also provides 4 examples showing that blood leukocyte markers, when used in combination with endometrial markers, can also increase the predictive value of the diagnostic test. The present invention also demonstrate that logistic regression models can also be used to combine endometrial as well as blood leukocyte markers for the development of a predictive model of endometriosis (Table 2) . In some cases, these models need to be adjusted with risk factors associated with endometriosis such as the length of the menstrual cycle, the duration of menstruation, pain (during intercourse, menstruation or in other circumstances) and age. In some instances, these factors were shown to increase the predictive value of the model . The present invention identifies several examples of marker combinations, which give rise to diagnostic methods yielding improved levels of sensibility and specificity. Indeed, the different marker combinations of the present invention may serve different clinical applications including screening, diagnosis, monitoring and prognosis of endometriosis. While the invention has been described in connection with specific embodiments thereof, it will be understood that it is capable of further modifications and this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains and as may be applied to the essential features hereinbefore set forth, and as follows in the scope of the appended claims. For example, other blood or endometrial markers, alone or in combination, might also be suitable to practice the method of the present invention, and are thus intended to be included in the present invention.

Claims: WHAT IS CLAIMED IS; 1. An endometrial or blood leukocyte marker for endometriosis characterized in that said marker is a surface antigen from endometrial or blood leukocytes. 2. The marker of claim 1, wherein the surface antigen from blood leukocytes is selected from the group consisting of CD3+, CD8+, CD13+, CD14+, CD20+, CD36+, CD44+, CD57+, CD69+, CD122+, CD3+CD5+, CD3+CD45RA-, CD3+CD56-, CD3+CD57-, CD3+CD69-, CD3+CD122-, CD3+HLADR+, CD3-CD5+, CD3-CD16+, CD3- CD44+, CD3-CD57+, CD3-CD69+, CD3-CD45RO+, CD3-CD4-, CD3-CD8-, CD3-CD45RA-, CD3-CD56-, CD4-CD13+ , CD4 - CD36+, CD4-CD69+, CD4-CD45R0+, CD4-CD45RA-, CD8-CD44+, CD8-CD44-, CD13+CD44+, CD13+HLADR+, CD13+CD16-, CD13- HLADR+, CD13-CD44-, CD14+HLADR+, CD14+CD44+, CD14+CD45R0+, CD14+CD16-, CD14+CD122-, CD14-HLADR+, CD14-CD44-, CD20+HLADR+, CD20+CD44-, CD20-CD44+, CD20- CD69+, CD20-HLADR+, CD20-CD44-, CD20-CD69-, CD36- HLADR+, CD56-CD69+, CD56-CD122+, CD56-CD69-, CD56- CD122-, CD57-CD44-, CD3+CD57-HLADR+, CD3 -CD4-CD44+ , CD3-CD56+CD16-, CD3 -CD56-CD122 - , CD3 -CD57-CD44- , CD14+CD20+CD44-, CD14+CD20-CD44+ , CD14+CD20-CD44- , CD14-CD13-HLADR+, Ratio CD13+/CD3+, Ratio CD13+/CD8+, Ratio CD14+/CD3+, Ratio CD14+/CD8+, CD3-CD57+CD44- , CD14-CD13-HLADR-, CD14-CD20+CD44 - , HLADR+ , CD3+CD8-, CD3+CD44-, CD3+HLADR-, CD3-HLADR+, CD3+CD16+, CD3- CD57-, CD4-HLADR+, CD4+CD45RA-, CD4+CD45RO+, CD13- CD16+, CD14-CD69-, CD20-HLADR-, CD57-CD44+ , CD3 - CD4+CD45RA-, CD3 -CD8-CD44+, CD3 -CD57-CD44+ , and CD14+CD20+CD44+. 3. The marker of claim 2 , wherein the surface antigen is chosen from CD3-CD57+CD44- , CD14-CD13- HLADR-, CD14-CD20+CD44-, HLADR+, CD3+CD8-, CD3+CD44-, CD3+HLADR-, CD3-HLADR+, CD3+CD16+, CD3-CD57-, CD4- HLADR+, CD4+CD45RA-, CD4+CD45RO+, CD13-CD16+, CD14- CD69-, CD20-HLADR-, CD57-CD44+ , CD3 -CD4+CD45RA- , CD3 - CD8-CD44+, CD3-CD57-CD44+, and CD14+CD20+CD44+ . 4. The marker of claim 1, wherein the surface antigen from endometrial leukocytes is selected from the group consisting of CD3+, CD4+, CD5+, CD8+, CD3+CD4+, CD3+CD4-, CD3-CD4-, CD3+CD8+, CD3+CD8-, CD3- CD8-, CD3+CD69+, CD3+CD122-, CD3+HLADR-, CD3-HLADR-, CD3+CD45RA+, CD3+CD45RA-, CD3-CD45RA-, CD3+CD45RO-, CD3+CD16-, CD3+CD56-, CD3+CD5+, CD3-CD5-, CD4+CD69-, CD4+CD45RA-, CD8+CD69-, CD8+HLADR-, CD8-HLADR-, CD13- CD122+, CD13-CD122-, CD20-CD5+, CD20-CD5-, CD56-CD16+, CD56-CD16-, ratio CD3+/CD45RO+, CD14+CD13-, CD3+CD20-, CD3-CD20-, CD3-CD4-CD45RA+, CD3 -CD4-CD45RA- , CD3+CD8+CD69-, CD3+CD8+HLADR- , CD3+CD8 -HLADR- , CD3 - CD8-HLADR-, CD14+CD13 -CD16b- , CD4+CD14-, CD4-CD14-, CD4+HLADR-, CD13-CD69+, CD13+CD45RO- , CD56-CD122-, CD3+CD69-, CD4+CD45RO-, CD56+CD122+, CD3 -CD56+CD16+, CD3-CD56+CD122+, CD14+CD13+, CD3+CD20-CD5+ , CD4- CD13+CD16+, CD69+, ratio CD13+/CD3+, CD3-CD20-CD5- , CD3+CD20-CD5-, CD4+CD13-, CD3+CD44-, CD56+, CD13- CD45RO+, CD56+CD69-, CD13-CD16+, CD56+CD122-, CD3+CD4- CD69+, CD4-CD13-CD16+, CD4 -CD13 -CD16- , CD14+CD13+CD16b+, CD4-CD69-, CD3-CD45RO+, CD4-CD69+, CD3-CD4-CD45RO+, CD3+HLADR+, CD3+CD45RO+, CD4+CD13+, CD13+HLADR+, CD14-HLADR+, CD56+HLADR+, CD56-CD44-, CD3-CD4+CD45RA-, CD3+CD8-HLADR+ , CD3 -CD8+HLADR- , CD3+CD56+CD16+, CD3+CD56-CD16+ , CD3+CD56-CD16- , CD3+CD56-CD44+, CD3+CD56-CD44- , CD3 -CD56-HLADR- , CD3+CD56-CD122+, CD3+CD56-CD122 - , CD4-CD16-, CD14+CD13+CD16b-, CD3-CD44-, CD16+, CD45RA+, CD45RO+, and CD13+. 5. The marker of claim 4, wherein the surface antigen is chosen from CD3+HLADR+, CD3+CD45RO+, CD4+CD13+, CD13+HLADR+, CD14-HLADR+, CD56+HLADR+, CD56-CD44-, CD3 -CD4+CD45RA- , CD3+CD8-HLADR+, CD3- CD8+HLADR-, CD3+CD56+CD16+ , CD3+CD56-CD16+ , CD3+CD56- CD16-, CD3+CD56-CD44+, CD3+CD56-CD44- , CD3 -CD56-HLADR- CD3+CD56-CD122+, CD3+CD56-CD122 - , CD4-CD16-, CD14+CD13+CD16b-, CD3-CD44-, CD16+, CD45RA+, CD45RO+, and CD13+. 6. A combination of markers for endometriosis comprising at least two different surface antigens from blood or endometrial leukocytes . 7. The combination of claim 6, wherein the surface antigens are selected from the group consisting of CD3+, CD4+, CD5+, CD8+, CD13+, CD14+, CD20+, CD36+, CD44+, CD56+, CD57+, CD69+, CD122+, HLADR+,CD16+, CD45RA+ , CD45R0+ , CD56-CD122+, CD3- CD8+HLADR-, CD3+CD4-CD69+ , CD3+CD4+, CD3+CD4-, CD3- CD4-, CD3+CD5+, CD3-CD5+, CD3-CD5-, CD3+CD8+, CD34-CD8- , CD3-CD8-, CD3+CD16+, CD3-CD16+, CD3+CD16-, CD3+CD20- CD3-CD20-, CD3+CD44-, CD3-CD44+, CD3-CD44-, CD3+CD45RA-, CD3-CD45RA-, CD3+CD45RA+, CD3+CD45RO+, CD3-CD45RO+, CD3+CD45RO-, CD3+CD56-, CD3-CD56-, CD3+CD57-, CD3-CD57+, CD3-CD57-, CD3+CD69-, CD3+CD69+, CD3-CD69+, CD3+CD122-, CD3+HLADR+, CD3+HLADR-, CD3- HLADR+, CD3-HLADR-, CD4+CD13+, CD4+CD13-, CD4-CD13+, CD4+CD14-, CD4-CD14-, CD4-CD16-, CD4-CD36+, CD4+CD45RA-, CD4-CD45RA-, CD4+CD45RO+, CD4+CD45R0-, CD4-CD45RO+, CD4+CD69-, CD4-CD69+, CD4-CD69-, CD4+HLADR-, CD4-HLADR+, CD8-CD44+, CD8-CD44-, CD8+CD69-, CD8+HLADR-, CD8-HLADR-, CD13+CD16-, CD13- CD16+, CD13+CD44+, CD13-CD44-, CD13+CD45RO- , CD13- CD45RO+, CD13-CD69+, CD13-CD122+, CD13-CD122-, CD13+HLADR+, CD13-HLADR+, CD14+CD13+, CD14+CD13-, CD14+CD16-, CD14+CD44+, CD14-CD44-, CD14+CD45RO+, CD14-CD69-, CD14+CD122-, CD14+HLADR+, CD14-HLADR+, CD20-CD5+, CD20-CD5-, CD20+CD44-, CD20-CD44+, CD20- CD44-, CD20-CD69+, CD20-CD69-, CD20+HLADR+, CD20- HLADR+, CD20-HLADR-, CD36-HLADR+, CD56-CD16+, CD56- CD16-, CD56-CD44-, CD56+CD69-, CD56-CD69+, CD56-CD69-, CD56+CD122+, CD56+CD122-, CD56-CD122-, CD56+HLADR+, CD57-CD44+, CD57-CD44-, CD3 -CD4 -CD44+ , CD3 -CD4+CD45RA- , CD3-CD4-CD45RA+, CD3 -CD4-CD45RA- , CD3 -CD4 -CD45RO+, CD3-CD8-CD44+, CD3+CD8+CD69- , CD3+CD8+HLADR- , CD3+CD8- HLADR+, CD3+CD8-HLADR-, CD3 -CD8-HLADR- , CD3+CD20-CD5+ , CD3+CD20-CD5-, CD3 -CD20-CD5- , CD3+CD56+CD16+, CD3+CD56-CD16+, CD3+CD56-CD16- , CD3 -CD56+CD16+ , CD3 - CD56+CD16-, CD3+CD56-CD44+, CD3+CD56-CD44 - , CD3+CD56- CD122+, CD3+CD56-CD122-, CD3 -CD56+CD122+ , CD3-CD56- CD122-, CD3-CD56-HLADR-, CD3 -CD57+CD44- , CD3-CD57- CD44+, CD3-CD57-CD44-, CD3+CD57-HLADR+, CD4- CD13+CD16+, CD4-CD13-CD16+, CD4-CD13 -CD16- , CD14+CD13+CD16b+, CD14+CD13+CD16b- , CD14+CD13 -CD16b- , CD14-CD13-HLADR+, CD14 -CD13 -HLADR- , CD14+CD20+CD44+, CD14+CD20+CD44-, CD14+CD20-CD44+ , CD14+CD20-CD44- , CD14-CD20+CD44-, ratio CD3+/CD45RO+, Ratio CD13+/CD3+, Ratio CD13+/CD8+, Ratio CD14+/CD3+, and Ratio CD14+/CD8+. 8. A combination of markers for endometriosis selected from the group consisting of: a) CD4+, CD8+CD69-, and CD13+CD45RO- ; b) CD4+, CD8+CD69-, CD56+CD122-, CD3+CD45RA- , and CD13+CD45RO-; c) CD4+, CD8+CD69-, CD13-CD122+, and CD13+CD45RO-; d) CD4+, CD8+CD69-, and CD14+CD13 -CD16b- ; e) CD3+CD16-, CD13+CD45RO- , CD3+,CD8+, and CD3+CD69+; f) CD3+, CD3+CD8+, CD13+CD45RO- , and CD3-CD20-; g) CD3+CD8+, CD13+CD45RO- , CD3+CD5+, CD3+CD122-, CD3-CD20-, and CD3+CD45RO-; h) CD3+CD8+, CD13+CD45RO-, CD3+CD5+, and CD3+CD122-; i) CD3+CD20-CD5-, CD4+CD13-, and CD56-CD122-; j) CD3+CD8+, CD13+CD45RO-, CD4+CD45RA-, and CD3+CD45RO-; k) CD3+, CD8+CD69-, CD3 -CD4-CD45RO+ , Ratio CD13+/CD3+, and CD13+CD45RO- ; 1) CD3+CD8+, CD13+CD45RO-, CD3-CD5-, and CD20- CD5+; m) CD8+, CD5+, CD3-CD20-, and CD3-HLADR-; n) CD3+CD8+, CD13+CD45RO- , and CD5+; o) CD4+, CD13-CD122+, CD8+CD69-, and CD3+CD45RA-; p) CD4+, CD8+CD69-, CD13-CD122+, CD3+CD45RA-, and CD13+CD45RO-; q) CD4+, CD8+CD69-, CD56-CD122-, CD3+CD45RA-, and CD13+CD45RO-; r) CD3+, CD4+, CD3+CD8+, CD13+CD45RO- , and CD3+CD5+; s) CD3+, CD4+, CD3+CD8+, and CD13+CD45RO- ; t) CD3+, CD3+CD8+, and CD13+CD45RO- ; u) CD3+, CD5+, CD3+CD5+, CD69+, and CD4-CD69+; v) CD3+ , CD3+CD84-, and CD13+CD45RO- ; w) CD3+, CD3+CD8+, and CD13+CD45RO- ; x) CD3+, CD3+CD8+, CD13+CD45RO- , CD3-CD20-, and CD56-CD16-; y) CD3+CD16-, CD3 -CD4-CD45RO+ , CD3+, CD8+, CD3+CD69+, and CD13+CD45RO- ; z) CD3+, CD3-CD45RO+, Ratio CD13+/CD3+, CD3+CD8+, and CD8+CD69-; aa) CD3+CD20-CD5-, CD4+CD13-, CD56-CD122-, and CD4+CD45RO-; bb) CD3+, CD3+CD8+, CD13+CD45RO- , and CD3-CD20-; cc) CD13+CD45RO-, CD4+CD45RA-, CD3+CD122-, and CD8+CD69-; dd) CD3+CD8+, CD13+CD45RO- , CD4+CD45RA-, CD3+CD45RO-, and CD3-CD5-; ee) CD3-CD5+, CD3-CD45RA-, CD3-CD44+, CD13+, and CD3-CD57-CD44-; ff) CD3-CD45RA-, CD20-CD44+, CD20-HLADR+, CD3- CD4-CD44+, and CD36-HLADR+; gg) CD3-CD45RA-, CD3-CD45RO+, and CD20-HLADR+; hh) CD57+, CD14+, CD3-CD69+, CD3+, CD4+, and CD3+CD8+HLADR-; ii) CD3-CD69+, CD3+, CD4+, and CD3+CD8+HLADR- ; jj) CD4-CD36+, CD3-CD69+, CD8+, CD13+, CD3+, CD16+, CD69+, and CD5+; kk) CD3-CD45RA-, CD3-CD45RO+, CD20-HLADR+, CD14+CD44+, CD8+, CD5+, CD3-CD20-, and CD3 - HLADR- ; 11) CD3+, CD3-CD5-, CD13+CD45RO- , and CD3-CD20-; mm) CD3+, CD3-CD5-, and CD13+CD45RO- ; nn) CD3+CD8+, CD13+CD45RO- , and CD3+CD5+; oo) CD3+, Length of menstruation, CD13+CD20-, and Pelvic pain; pp) CD14+CD44+, CD57+, CD3-CD45RA-, and CD14+; qq) CD14+, CD57+, CD3+CD69+, CD3+HLADR+, and CD3-CD45RA-; rr) CD14+, CD57+, CD3+HLADR-, CD3-CD45RA-, Pelvic pain, and Length of menstruation; and ss) CD14+, CD57+, CD3+CD69+, CD3+HLADR-, CD3 - CD45RA-, Pelvic pain, and Age. 9. A diagnostic method for the detection of endometriosis in a patient sample, said method comprising the step of detecting at least one marker of any one of claims 1, 2, 3, 4, and 5, whereby detection of said specific marker is indicative of endometriosis . 10. A diagnostic method for the detection of endometriosis in a patient sample, said method comprising the step of detecting at least two different surface antigens from blood or endometrial leukocytes, whereby detection of said at least two different surface antigens is indicative of endometriosis . 11. A diagnostic method for the detection of endometriosis in a patient sample, said method comprising the step of detecting a combination for endometriosis as defined in any one of claims 6, 7, and 8, whereby detection of said combination is indicative of endometriosis. 12. A diagnostic kit for the detection of endometriosis, said kit comprising an antibody specific for a marker as defined in any one of claims 1 , 2 , 3 , 4 , and 5. 13. A diagnostic kit for the detection of endometriosis, said kit comprising at least two different antibodies, each specific for different surface antigens as defined in the combination of claim 6, 7 or 8. 14. The diagnostic kit of claim 13, wherein said specific marker combination is selected from the group consisting of: a) CD4+, CD8+CD69-, and CD13+CD45RO- ; b) CD4+, CD8+CD69-, CD56+CD122-, CD3+CD45RA-, and CD13+CD45RO-; c) CD4+, CD8+CD69-, CD13-CD122+, and CD13+CD45RO-; d) CD4+, CD8+CD69-, and CD14+CD13-CD16b- ; e) CD3+CD16-, CD13+CD45RO- , CD3+,CD8+, and CD3+CD69+; f) CD3+, CD3+CD8+, CD13+CD45RO- , and CD3-CD20- g) CD3+CD8+, CD13+CD45RO- , CD3+CD5+, CD3+CD122-, CD3-CD20-, and CD3+CD45RO-; h) CD3+CD8+, CD13+CD45RO-, CD3+CD5+, and CD3+CD122-; i) CD3+CD20-CD5-, CD4+CD13-, and CD56-CD122-; j) CD3+CD8+, CD13+CD45RO-, CD4+CD45RA-, and CD3+CD45RO-; k) CD3+, CD8+CD69-, CD3 -CD4-CD45RO+ , Ratio CD13+/CD3+, and CD13+CD45RO- ; 1) CD3+CD8+, CD13+CD45RO-, CD3-CD5-, and CD20- CD5+; m) CD8+, CD5+, CD3-CD20-, and CD3-HLADR-; n) CD3+CD8+, CD13+CD45RO- , and CD5+; o) CD4+, CD13-CD122+, CD8+CD69-, and CD3+CD45RA-; p) CD4+, CD8+CD69-, CD13-CD122+, CD3+CD45RA-, and CD13+CD45RO-; q) CD4+, CD8+CD69-, CD56-CD122-, CD3+CD45RA-, and CD13+CD45RO-; r) CD3+, CD4+, CD3+CD8+, CD13+CD45RO- , and CD3+CD5+; s) CD3+, CD4+, CD3+CD8+, and CD13+CD45RO- ; t) CD3+, CD3+CD8+, and CD13+CD45RO- ; u) CD3+, CD5+, CD3+CD5+, CD69+, and CD4-CD69+; v) CD3+, CD3+CD8+, and CD13+CD45RO- ; w) CD3+, CD3+CD8+, and CD13+CD45RO- ; x) CD3+, CD3+CD8+, CD13+CD45RO- , CD3-CD20-, and CD56-CD16-; y) CD3+CD16-, CD3 -CD4-CD45RO+ , CD3+, CD8+, CD3+CD69+, and CD13+CD45RO- ; z) CD3+, CD3-CD45RO+, Ratio CD13+/CD3+, CD3+CD8+, and CD8+CD69-; aa) CD3+CD20-CD5-, CD4+CD13-, CD56-CD122-, and CD4+CD45R0-; bb) CD3+, CD3+CD8+, CD13+CD45RO- , and CD3-CD20-; cc) CD13+CD45RO-, CD4+CD45RA-, CD3+CD122-, and CD8+CD69-; dd) CD3+CD8+, CD13+CD45RO- , CD4+CD45RA-, CD3+CD45RO-, and CD3-CD5-; ee) CD3-CD5+, CD3-CD45RA-, CD3-CD44+, CD13+, and CD3-CD57-CD44-; ff) CD3-CD45RA-, CD20-CD44+, CD20-HLADR+, CD3 - CD4-CD44+, and CD36-HLADR+; gg) CD3-CD45RA-, CD3-CD45RO+, and CD20-HLADR+ ; hh) CD57+, CD14+, CD3-CD69+, CD3+, CD4+, and CD3+CD8+HLADR-; ii) CD3-CD69+, CD3+, CD4+, and CD3+CD8+HLADR- ; jj) CD4-CD36+, CD3-CD69+, CD8+, CD13+, CD3+, CD16+, CD69+, and CD5+; kk) CD3-CD45RA-, CD3-CD45RO+, CD20-HLADR+, CD14+CD44+, CD8+, CD5+, CD3-CD20-, and CD3- HLADR- ; 11) CD3+, CD3-CD5-, CD13+CD45RO- , and CD3-CD20-; mm) CD3+, CD3-CD5-, and CD13+CD45RO- ; nn) CD3+CD8+, CD13+CD45RO- , and CD3+CD5+; oo) CD3+, Length of menstruation, CD13+CD20-, and Pelvic pain; pp) CD14+CD44+, CD57+, CD3-CD45RA-, and CD14+; qq) CD14+, CD57+, CD3+CD69+, CD3+HLADR+, and CD3-CD45RA-; rr) CD14+, CD57+, CD3+HLADR-, CD3-CD45RA-, Pelvic pain, and Length of menstruation; and ss) CD14+, CD57+, CD3+CD69+, CD3+HLADR-, CD3- CD45RA-, Pelvic pain, and Age. 15. Use of a marker as defined in any one of claims 1, 2, 3, 4, and 5 for identifying a woman suffering or likely to suffer from endometriosis. 16. Use of a combination as defined in any one of claims 6, 7, and 8 for identifying a woman suffering or likely to suffer from endometriosis. 17. Use of a diagnostic kit as defined in any one of claims 12, 13, and 14 for identifying a woman suffering or likely to suffer from endometriosis.

Inventors: GAGNE DANIELE, GOSSELIN DIANE, HUGO PATRICE, MIRON PIERRE, GAGNE, DANIELE, GOSSELIN, DIANE, HUGO, PATRICE, MIRON, PIERRE
Applicants: GAGNE DANIELE, GOSSELIN DIANE, HUGO PATRICE, MIRON PIERRE, PROCREA BIOSCIENCES INC, GAGNE, DANIELE, GOSSELIN, DIANE, HUGO, PATRICE, MIRON, PIERRE, PROCREA BIOSCIENCES INC.
Application year: 2000
Main classification: G01N33
IPC codes: G01N33