bluuebunny/crossref_metadata_2025 huggingface dataset:
The significance of activation markers on CD8 lymphocytes in human immunodeficiency syndrome: staging and prognostic value
Authors: LEVACHER M , HULSTAERT F , TALLET S , ULLERY S , POCIDALO J J , BACH B A
Published: July 2010

The objective of this prospective cohort study was to evaluate the expression of activation markers on CD8 lymphocytes at various clinical stages of HIV infection and to determine the value of these markers in identifying patients likely to have rapidly progressive disease. One hundred and three HIV+ patients, divided into four disease stages, and 34 seronegative controls were evaluated at study entry using flow cytometric immunophenotyping. The HIV patients were followed clinically for disease progression during the following 2 years. CD8 cell numbers and percentage of lymphocytes are increased after HIV infection. Expression of the CD38, HLA-DR and CD57 markers on CD8 cells was significantly increased in asymptomatic HIV-infected patients when compared with controls, as was the CDS cell population which did not coexpress Leu-8. These activation markers were observed to be further increased in patient groups with more clinically advanced infection. The percentage of CD38 on CD8 cells emerged not only as a discriminator of disease severity, but was a strong predictor of progression in asymptomatic, lymphadenopathy and ARC patients. Given the utility of activation markers on CDS lymphocytes in staging disease and predicting clinical outcome, the measurement of these parameters should be considered in the monitoring and management of HIV patients.
Use of recombinant epitopes to study the heterogeneous nature of the autoantibodies against thyroid peroxidase in autoimmune thyroid disease
Authors: ZANELLI E , HENRY M , MALTHIERY Y
Published: July 2010

Microsomal antigen is often recognized by the sera from patients with autoimmune thyroid disease (AITD). Human thyroid peroxidase (hTPO) is the main component of this antigen. In a previous study, we expressed hTPO cDNA as fusion proteins in prokaryotic vector; we thereby defined seven antigenic peptides by using two rabbit polyclonal anti-hTPO antibodies. In the present study we used the seven epitopes and three widened peptides to define the reactivity pattern of 61 sera from patients with AITD. Thirty-eight of them reacted against at least one of the seven hTPO-restricted epitopes; 14 were negative against the seven determinants but recognized one or two of the extended peptides. Thus, the antibody response against hTPO appeared to be highly heterogeneous in AITD patient sera. Moreover, we demonstrated that the immunodetection of the hTPO on Western blotting with deoxycholate solubilized microsomes can be perfectly correlated with the recognition of one of the epitopes in the region 554–735.
Peritoneal macrophages during peritonitis. Phenotypic studies
Authors: HART P H , JONES C A , FINLAY-JONES J J
Published: July 2010

The expression of a range of surface molecules/receptors that are important in the host response to infection and foreign antigens was examined using peritoneal macrophages isolated from patients on continuous ambulatory peritoneal dialysis (CAPD) with peritonitis. The macrophage phenolypic profile was compared with that of normal peripheral blood monocytes. Consistently there was increased expression by macrophages of CD14, ICAM-1 (CD54). FcγRI (CD64). FcγRII (CDw32), FcγRIII (CD 16), transferrin receptors (CD71)and tissue factor. Increased expression of MHC class II was marginally significant. There was no detectable expression of either the p55 (CD25) or p70 chains of the IL-2 receptor. The expression of the complement receptors. CRI (CD35) and CR3 (CD11b. CDI8), was reduced. The activity of well-known inflammatory cytokines, rather than uraemic molecules, can account for the phenotypic profile of these extravasated peritoneal macrophages. The results of this study indicate that peritoneal macrophages from CAPD patients with peritonitis display a phenotype consistent with them being in vivo-derived inflammatory macrophages, and that they arc appropriate for use in studies of anti-inflammatory agents.
Kinetics of cytokine secretion by mononuclear cells of the blood from rheumatoid arthritis patients are different from those of healthy controls
Authors: RUSCHEN S , STELLBERG W , WARNATZ H
Published: July 2010

Mononuclear cells from peripheral blood (PBMC) of rheumatoid arthritis (RA) patients and healthy controls were incubated with α-CD3. Cytokine secretion from 2h to 72 h of incubation was measured by ELISA, There were no significant differences in secretion of T cell derived IL-2 and IL-4 in cultures from RA patients and controls. The macrophage-derived cytokines, IL-1β and tumour-necrosis factor-α (TNF-α) were secreted with a steep increase of concentration during the first 16 h of incubation by PBMC from RA patients. PBMC from healthy controls secreted both cytokines at a constantly rising rate with a maximum for TNF-α at 48 h and for IL-1β at 72 h. Interferon-γ (IFN-γ) is secreted in significantly reduced concentrations by PBMC from untreated RA patients compared with controls. Gold-salt treatment led to a slightly delayed and enhanced secretion of TNF-α and IL-1β, an enhanced secretion of IL-2 and a restored secretion of IFN-γ.
T cell receptor Vβ repertoire in HIV-infected individuals: lack of evidence for selective Vβ deletion
Authors: BOYER V , SMITH L R , FERRE F , PEZZOLI P , TRAUGER R J , JENSEN F C , CARLO D J
Published: July 2010

The gradual decline of CD4+ T lymphocytes in HIV-infected individuals culminates in the lethal immunosuppression of AIDS. The mechanism of CD4+ T cell loss is currently unknown, but has recently been suggested to occur as a result of an HIV-encoded superantigen which facilitates a selective deletion of T cells expressing specific Vβ genes. To verify and extend such observations, peripheral blood leucocytes (PBL) from 15 CD4+ individuals, 10 of which had very low CD4 T cell counts (< 200/mm3), were analysed for T cell receptor (TCR) V/β gene expression. In contrast to a recent study, the results presented here fail to provide evidence that selective loss of Vβ-bearing T cells occurs in HIV+ individuals. Furthermore, when PBL from HIV+ individuals were stimulated with Staphylococcal enterotoxin B (SEB), T cells expressing Vβ subfamilies known to engage this superantigen were expanded, indicating that such cells were not deleted and were responsive to stimulation by a bacterial superantigen. Collectively, these data suggest that CD4 loss in HIV patients docs not occur in a Vβ-seleetive, superantigen-mediated fashion.
Pigeon breeders’ lung; IgG subclasses to pigeon intestinal mucin and IgA antigens
Authors: TODD A , COAN R , ALLEN A
Published: July 2010

Sera from all symptomatics with pigeon breeders’ lung and many non-symptomatic pigeon breeders react specifically with the pigeon antigens and in particular pigeon intestinal secretions and isolated pigeon intestinal mucin and IgA. In sera from all 82 precipitin-positive pigeon breeders investigated, IgGl reacted with both IgA and mucin, while IgG2 reacted predominantly with mucin only. Sera from all 32 symptomatic patients with pigeon breeders’ lung demonstrated an IgG3 reaction with mucin, but only three sera showed reactivity against IgA and this was very weak. In contrast, only three out of 27 pigeon breeders with pulmonary pathology other than pigeon breeders’ lung and only three out of 23 sera from healthy pigeon breeders showed detectable IgG3 reactivity with mucin. IgG4 from eight sera only reacted with mucin (n= 7) or IgA (n= 1) and showed no consistent pattern between subject groups. Precipitin-negative sera from 30 volunteers with no previous pigeon exposure showed no IgG subclass reactivity with either the pigeon intestinal secretions or the isolated mucin and IgA antigens. These results show specific IgG2 and IgG3 subclass reactivity to pigeon intestinal mucin antigen in precipitin-positive sera from pigeon breeders. Further IgG3 subclass reactivity specific to pigeon mucin is strongly associated with sera from patients with the active pigeon breeders’ lung disease.
Exposure to complement-bearing immune complexes enhances thein vitrosequestration of erythrocytes from young but not elderly donors
Authors: SHAPIRO S , PILAR T , GERSHON H
Published: July 2010

Complement and immunoglobulin have each been claimed to be the major opsonins responsible for sequestration of the effete erythrocyte. Binding of immune complexes to the erythrocyte via CRI (CD35) provides a model for studying the effects of increments in metnbrane-bound complemenl and immunoglobulin on the sequestration of the erythrocyte (‘innocent bystander’ sequestration). It is possible that C3b-bearing immune complexes (IC C3b) bound to erythrocyte CRI contribute to the levels of immunoglobulin and complement fragments detectable on the human erythrocyte. We have, therefore, compared the capacity of erythrocytes from young and elderly donots to bind IC C3b and the effect of this binding on in vitro sequestration. Erythrocytes from young donors exposed to IC-C3b bind these complexes, as attested by an increment in membrane-bound C3, and undergo ‘innocent bystander’ sequestration. However., when density-separated erythrocytes are so exposed, it is only the low density (young) erythrocytes from young donors which are susceptible to ‘innocent bystander’ sequestration. High density (old) erythrocytes from young donors and ail erythrocytes from elderly donors show initially high in vitro sequestration and are resistant to the “inniwent bystander’ effect. (Those erythrocytes which show initially high in vitro sequestration are referred to collectively as in situ aged’ erythrocytes.) There is a great similarity between the mechanisms of sequestration of in situ aged’ and “innocent bystander” erythrocytes in that they are both inhibited by the integrin binding peptide arginine-glycine-aspartic acid (RGD) and the β-galactosyl sugar n-acetyl-galactosamine, and unaffected by the Fe-γ binding protein, Protein-G. Complement is the major opsonin in Innocent bystander’ sequestration since this sequestration occurs whether the isotype of the antibody in the immune complex is IgM or IgG. and Protein-G, which inhibits IgG-dependent erythrophagocytosis. has no effect on ‘innocent bystander’ sequestration. The present studies demonstrate that in vitro sequestration of ‘in situ aged’ erythrocytes is similar to complement-dependent ‘innocent bystander’ sequestration, thus supporting the contention that complement recognition is eardinal in sequestration of ‘in situ aged’ erythrocytes.
IL-1β and IL-3-like activity in preterm infants
Authors: BESSLER H , SIROTA L , NOTTI I , DULITZKY F , DJALDETTI M
Published: July 2010

The capacity of peripheral blood mononuclear cells (PBMC) of pretert ntieonaies to release IL-1β and IL-3-like activity (IL-3-LA) has been investigaied. In the present study it was found that this capacity is significantly lower than that of their mothers and or eontrol adults. In addition, the results showed that preterm serum has a lower stitnulatory effect on IL-1β production and an inhibitory effect on IL-3-LA secretion by PBMC of adult controls, in comparison with maternal and adull sera. These findings suggest an additional Teedback mechanism for control of hacmatopoiesis in premature neonates. It is possible that the lower production of IL-1β and IL-3-VLA may be involved in the increased susceptibility to infections of preterm newboms.
Renal allograft rejection: expression and function of VCAM-1 on tubular epithelial cells
Authors: LIN Y , KIRBY J A , BROWELL D A , MORLEY A R , SHENTON B K , PROUD G , TAYLOR R M R
Published: July 2010

The interaction between vascular cell adhesion molecule-1 (VCAM-1) and very late antigen-4 (VLA-4) is known to play an important role in stabilizing the adhesion of lymphocytes to endothelial cells. Such cellular adhesion is crucial to many immunological processes including lymphocyte-mediated cell lysis. In this study the expression of VCAM-I on renal tubular epithelial cells is demonstrated on biopsy sections recovered during acute renal allograft rejection. Experiments performed using epithelial cells cultured from renal tubules show that VCAM-1 is up-regulated by addition of the inflammatory cytokines tumour necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ). Combination of TNF-α and IFN-γ synergized to induce high levels of VCAM-1 expression. Further experiments demonstrated that the cytokines produced by activated lymphocytes in mixed leucocyte culture also up-regulate expression of VCAM-1. Assays of the adhesion of lymphoid cells to cultured renal epithelial cells showed that cytokine pretreatment of the renal cells enhanced the binding of lymphoid cells. The proportion of bound lymphoid cells was significantly reduced by addition of an antibody capable of blocking the interaction of VCAM-1 with VLA-4. This result indicated that the VCAM-1 induced on renal epithelial cells by inflammatory cytokines is functionally capable of binding VLA-4, thereby enhancing the adhesion of potentially graft-damaging lymphoid cells.
Measurement of antiphospholipid antibody by ELISA using purified β2-glycoprotein I in preeclampsia
Authors: YAMAMOTO T , YOSHIMURA S , GESHI Y , SASAMORI Y , OKINAGA S , KOBAYASHI T , MORI H
Published: July 2010

It has been reported that the binding of some antiphospholipid antibodies (APA) to phospholipids requires the presence of β2-glycoprotein 1 (β2-GPI). Using a new ELISA, in which well coated phospholipids were treated with a constant amount of purified β2-GPI. we tried to detect the presence of APA which binds to phospholipid/β2-GPI complex or to phospholipids such as cardiolipin (CA) and phosphatidylserine (PS) in preeclampsia. and to check for clinical abnormalities in antibodypositive cases. Serum samples were taken from 43 cases of preeclampsia. including 26 cases of the severe type, and 47 normal pregnant women. Positive rates of anticardiolipin antibody (ACA) by ELISA using CA/β2-GPI complex in mild, severe and total preeclampsia were 20·0%, 17·4% and 18·4% respectively. No antibody-positive cases were found in normal pregnancies. ACA was detected much more frequently when cardiolipin/β2-GPI complex was used in ELISA compared with ELISA without β2-GPI. Positive rales of antiphosphatidylserine antibody (APSA) in mild, severe and total preeclampsia were 5·9%, 11·5% and 9·3% respectively. APSA was also detected much more frequently when the phosphatidylserine/β2-GPI complex was used in ELISA. The frequency of intraulerine growth retardation (IUGR) in the ACA-positive subjects was higher than that of ACA negatives. We suggest that the ACA and APSA which bind to phospholipid/β2-GPI complex are detectable in preeclampsia, and that these antiphospholipid antibodies are related to fetal growth.
Limiting dilution analysis of the allo-MHC anti-paternal cytotoxic T cell response II: recurrent spontaneous abortion and the effect of immunotherapy
Authors: MANYONDA I T , PEREIRA R S , PEARCE J M , SHARROCK C E M
Published: July 2010

Using limiting dilution analysis (LDA) we determined anti-paternal cytotoxic T lymphocyte precursor (CTLp) frequencies in the peripheral blood of 10 women with unexplained recurrent spontaneous abortion (RSA) before and after immunization with paternal lymphocytes. The women and their partners were HLA tissue-typed and none of the women had anti-paternal cytotoxic antibodies (APCA) before immunization. All other known causes of RSA were excluded. All 10 women were found to have high frequencies of specific anti-paternal cytotoxic T cells before immunization (range I 1/1030 to 1/9574). Splitwell analysis showed that these cytotoxic cells were specific to paternal MHC antigens. These frequencies rose significantly following immunization (range 1/683 to 1/4652). The cytotoxic T lymphocyte frequencies against an HLA-mismatched third party varied from woman to woman, but were not affected by the immunization. The LDA data conformed lo single-hit kinetics, indicating that only cytotoxic T ceils were limiting in the assay. Our data are in sharp contrast to the previously held view that women with RSA may be hyporesponsive to paternal MHC antigens. Immunizing such women with paternal leucocytes further sensitizes them. These findings cannot be reconciled with a favourable outcome in the treatment of RSA with immunotherapy. We would argue that this treatment is al best of unproven value, and may even be harmful. Thai these women may sometimes have successful pregnancies following immunotherapy testifies to the effectiveness of the classical MHC antigen-deficient trophoblast as an immunological barrier between mother and fetus.
Molecular basis for antibody cross-reactivity between the hepatitis C virus core protein and the host-derived GOR protein
Authors: ZHANG Z X , CHEN M , WALLHAGEN K , TROJNAR J , MAGNIUS L O , WAHRKN B , SÄLLBERG M
Published: July 2010

The presence of antibodies reactive to a recently cloned host-derived antigen GOR is highly correlated with the presence of antibodies to the hepatitis C virus (HCV), We explored the molecular basis for this observation, and address the following question: are antibodies reactive with GOR19-27 (QKAKSNPNR) a result of a cross-reactivity triggered by the antigenic region at residues 9-17 of HCV core (RKTKRNTNR)? We compared the relative antibody avidity between antibodies reactive to both regions, and determined the residues essential for antibody binding using substitution peptide analogues. Of 96 sera assayed, 60 were found positive for anti-HCV, and of these 55 were found to react with HCV core. Twenty-nine sera were found reactive to the GOR peptide, and these were all reactive to HCV core. In most cases the relative antibody avidity of antibodies reactive to GOR was higher for the HCV core peptide. In 21 of the GOR-reactive sera we were able to determine the essential residues for antibody binding. The essential residues in > 50% of all tested sera coincided with the well conserved residues Lys10, Lys12, Asn14, and Asn16. Also, reactivity to GOR was not related to any certain serotype of antibodies to HCV. Taken together, these findings explain at the molecular level the observed cross-reactivity between these two proteins, since sequence homology per se is not evidence for cross-reactivity.
HLA expression in hepatocellular carcinoma cell lines
Authors: WADEE A A , PATERSON A , COPLAN K A , REDDY S G
Published: July 2010

The present study undertook to investigate the biological significance of human leucocyte antigen expression in hepatocellular carcinoma and to elucidate the role of potential modulating agents on human leucocyte antigen expression. These studies used several hepatic tumour-derived cell lines as in vitro model systems. The cell lines included PLC/PRF/5 (Alexander cell line), Hep3B. HepG2, TONG PHC, HA22T/VGH, HA59T/VGH and Mahlavu, The cell lines K562 and Raji were used as negative and positive controls, respectively. K562, a B tymphoid-derived cell line, was shown to express negligible amounts of human leucocyte anligens, while Raji, an erythromyeloid-derived cell line, expressed both class I and class II human leucocyte anligens as well as their respective invariant chains, β2-niicroglobulin and Ii. Using an ELISA, experiments performed on these cell lines confirmed the natural expression of class I and class II antigens by the HA22T/VGH and HA59T/VGH cell lines, whereas PLC/PRF/5 displayed class II surface antigens only. The effects of modulating agents such as interferon-gamma sodium bulyrate and clofazimine on human leucocyte antigen expression were investigated using the HA22T/VGH, HA59T/VGH and TONG PHC cell lines. These agents increased class 1 and class II human leucocyte antigen expression on HA22T/VGH and TONG PHC cells, but had no effect on the HA59T/VGH cell line. The results suggest a potential use for these agents as modulators of human leucocyte antigen expression by human heptocellular cell lines.
Study of IL-2 receptor expression after chemoimmunotherapy in patients treated for metastatic malignant melanoma
Authors: MOUAWAD R , ICHEN M , RIXE O , BENHAMMOUDA A , VUILLEMIN E , WEIL M , KHAYAT D , SOUBRANE C
Published: July 2010

Using flow cytometry, cellular IL-2 receptors were studied before and following chemoimmunotherapy combination in 20 patients with metastatic malignant melanoma (MMM). Patients received cisplatin (100 mg m2) at days 1 and 28, recombinant IL-2 by continuous infusion from days 3 to 6, 17 to 21, 31 to 34, and 45 to 49. Interferon-alpha (IFN-α) was given subcutaneously three times weekly. In terms of clinical response, we observed 55% objective response (complete: 15%), When pretreatment blood samples were compared with those of healthy donors, we did not observe any change in low (α chain) and high affinity receptor (α+β) expression. In contrast, intermediate affinity p75 (β chain) expression was decreased significantly (P ≤ 0.0001) in MMM patients. During treatment, we found a dramatic increase of β chain as well as high affinity (α+β) expression in responding patients, as soon as IL-2 therapy began. Furthermore, the increase of β chain expression was limited to natural killer (NK) cells (CD56+). In non-responding patients, on the other hand, increase of both receptors was seen only at day 31. These data suggest the involvement of β chain expression in the mechanism of cell activation after chemoimmunotherapy. Moreover, this early 3 chain expression is correlated with the clinical response to chemoimmunotherapy.
Anti-lymphocyte antibodies in plasma of HIV-1–infected patients preferentially react with MHC class Il-negative T cells and are linked to antibodies against gp41
Authors: MÜLLER C , KUKEL S , SCHNEWEIS K E , BAUER R
Published: July 2010

It has previously been shown that HIV-infected patients develop anti-lymphocyte antibodies. The relationship between anti-lymphocyte antibodies and antibodies against different viral antigens is unknown, and it remains controversial whether some lymphocyte subpopulations are targeted preferentially. We have set out using three-colour flow cytometry to measure antibodies against different lymphocyte subsets. Staining with anti-human immunoglobulin and two MoAbs was performed to characterize the immunoglobulin toad of different lymphocyte subsets. Comparison was done between patients'antibody reactivity against HlV-1 antigens and anti-lymphocyte antibodies. We were able to demonstrate the presence of anti-lymphocyte antibodies in approximately 75% of the HIV-infected patients (n = 78) (healthy controls were all negative). MHC class Il-negative T cells showed a stronger reaction with anti-lymphocyte antibodies than B cells or MHC class Il-positive T cells. Patients with antibodies against CD4 lymphocytes showed a significantly higher antibody reaction with the retroviral antigen gp41 than patients without these antibodies. An association between anti-lymphocyte antibodies and antibody reactivity against other HIV-1 antigens was not noticed. In conclusion, anti-lymphocyte antibodies in HIV-1-infected patients show a preferential reactivity with T cells which lack expression of MHC class II molecules. There is an increased antibody reactivity against gp41 in patients with anti-CD4+ T cell antibodies. The association hints at a specific origin of anti-lymphocyte antibodies in HIV-1-infected patients due to cross-reactivity with viral epitopes or network phenomena. These anti-CD4 cell antibodies could be of interest in the clinical course of HIV infection.
Induction of multinucleated giant cells from rheumatoid arthritis (RA) synovial adherent cells by anti-DR antibody
Authors: MATSUYAMA T , NAKASHIMA N , MATSUDA T , NAKAMURA H , UCHIDA S , ABE T
Published: July 2010

To determine the effects of signalling through the DR molecule on synoviocytes from RA patients, the synovial adherent cells were incubated with anti-DR antibodies. After 24 h incubation, we found the formation of multinucleated giant cells in that culture. These multinucleated giant cells showed characteristics of monocyte-macrophage lineage cells and precursor of osteoclasts. Cyclohexamide inhibited the formation of multinucleated giant cells, but not the aggregation of synovial cells, suggesting that newly synthesized proteins are associated with the cell fusion. These results revealed a new mechanism in multinucleated giant cell formation.
Isolation and characterization of migratory human skin dendritic cells
Authors: RICHTERS C D , HOEKSTRA M J , VAN BAARE J , DU PONT J S , HOEFSMIT E C M , KAMPERDIJK E W A
Published: July 2010

A method is described to isolate and characterize human skin dendritic cells (DC). This method is based on the migratory capacities of these cells. The cells migrated “spontaneously” out of split-skin explants into the medium during a 24-h culture period and contained up to 75% CD1a+ cells. After removal of co-migrated T cells and macrophages, the highly enriched (>95% CD1a*) DC showed potent allo-antigen-presenting capacities. About 25% of the CD1a+ cells were also positive for the dermal DC marker CD1b, whereas only 15-20% of the cells contained Birbeck granules, the characteristic cell organelle of the epidermal Langerhans cell. Before culture. CD1a* DC were observed on cryostat sections not only in the epidermis but also in the dermis. After culture, the number of CD1a+ cells in both epidermis and dermis had decreased. Not all the cells had migrated during the culture period; some CD1a+ cells could still be detected in the epidermis and dermis after culture. Thus, using this method, potent allo-stimulating CD1a+ cells, migrating from both epidermis and dermis can be obtained without the use of enzymes.
Common variable immunodeficiency (CVID) and MxA-protein expression in blood leucocytes
Authors: RUMP J A , JAKSCHIESS D , WALKER U , SCHLESIER M , VON WUSSOW P , PETER H H
Published: July 2010

The underlying immunopathogenic mechanism of CVID has been suspected to involve a chronic viral infection or an autoimmune condition. However, formal proof of viral infection is lacking. Measurement of MxA-protein in leucocyte lysates is a sensitive test for evaluating the activation of the host's interferon system. Both viral infections and autoimmune diseases such as systemic lupus erythematosus (SLE) strongly induce MxA-protein in peripheral leucocytes. We therefore examined 15 patients with longlasting hypogammaglobulinaemia for MxA-protein induction in vivo: 13 patients suffered from CVID, one from hyper-IgM syndrome, and one patient had chronic B lymphocytic leukaemia associated with immunoglobulin deficiency and chronic papilloma virus infection (condylomata accuminata). Only the latter patient exhibited a strong MxA-protein expression; two CVID patients were borderline positive, and the remaining 12 patients including the hyper-IgM syndrome were MxA-protein-negative. There was no relationship between MxA expression and low CD4/CD8 ratios or increased CD8/CD57+ T cell counts, although both conditions are often observed in CVID as well as in chronic viral infections. When exposed in vitro to interferon-alpha (IFN-α), peripheral blood leucocytes of four MxA-negative patients were capable of producing normal amounts of MxA-protein. Taken together, these results argue against a viral or autoimmune pathogenesis of CVID.
Technical Report: results of immunological tests on faecal extracts are likely to be extremely misleading
Authors: Ferguson A , Humphreys K A , Croft N M
Published: July 2010

Clinical investigation of gut immunity is difficult because of the need to study intestinal tissues or secretions directly. Others have reported that immunoglobulins, antibodies and cytokines can be detected in saline extracts of faeces. We have assessed the validity of this approach by measuring immunoglobulins, albumin, α1-antitrypsin and isotype-specific antibodies in matched samples of faeces and whole gut lavage fluid. Results were compared as estimated output per day, and by using haemoglobin as a common reference substance. Samples were obtained from 10 patients with active inflammatory bowel disease and 10 with other benign GI diseases. For immunoglobulins, albumin and antibodies, the amount detected in faeces varied from < 0.01% to 35.5% (based on estimated daily output) and < 0.01 % to 18.5% (based on haemoglobin) of the amount known to be produced in the gut from results of assays on whole gut lavage fluid (WGLF); there were significantly higher rates of recovery in faecal specimens from patients with active gut inflammation than from other patients. Detection rates and titres of specific antibody in faeces were even lower than those for immunoreactive IgA. These data indicate that immunological tests on saline extracts of faeces do not represent the true status of the gut humoral immune system, and such studies should be strongly discouraged.
Differing antibody IgG isotypes in the polar forms of leprosy and cutaneous leishmaniasis characterized by antigen-specific T cell anergy
Authors: ULRICH M , RODRIGUEZ V , CENTENO M , CONVIT J
Published: July 2010

Leprosy and American cutaneous leishmaniasis are tropical diseases which present a spectrum of clinical and immunological manifestations. Lepromatous leprosy and diffuse cutaneous leishmaniasis are the severe, progressive polar forms of disease characterized by persistent T cell anergy. Relative concentrations of antibodies belonging to the four IgG isotypes have been determined in these forms of disease as well as active visceral leishmaniasis, which presents transitory T cell anergy. Leishmania-specific lgG4 antibodies predominated in 19/20 sera from patients with diffuse cutaneous leishmaniasis, and IgG1 antibodies predominated in 9/10 cases of untreated visceral leishmaniasis. The predominant IgG isotype of Mycobacterium leprae-specific antibodies in untreated lepromatous leprosy was remarkably variable (IgG1, igG2, IgG3 and IgG4 in 8, 6, 2 and 1 sera, respectively). Differing IgG antibody isotypes have been associated with distinct CD4+ T cell helper subpopulations and their characteristic lymphokine profiles in several pathologies. These results suggest that T cell anergy in chronic intracellular infections may be associated with as yet undefined mechanisms which modulate reported T helper cell lymphokine isotype relationships.
Antibodies to small ribonucleoprotein and to 73-kD heat shock protein: two distinct markers of mixed connective tissue disease
Authors: APPELBOOM T , KAHN M F , MAIRESSE N
Published: July 2010

We set out to discover whether antibodies to small ribonucleoprotein antigens (RNP) and to 73-kD heat shock protein (hsp 73) which have been proposed as markers of mixed connective tissue disease (MCTD) recognize different epitopes. MCTD serum was immunoadsorbed on hsp 73-coupled Sepharose and the affinity retained, and non-retained fractions were checked by immunoblotting for recognition of either purified bovine hsp 73 or calf thymus extract RNPs. The hsp 73 affinity-bound serum fraction recognized hsp 73 but not RNP antigens, the reverse being true for the non-retained fraction. We conclude that anti-hsp 73 and anti-RNPs are distinct markers of MCTD.
Interactions and molecular structure of cardiolipin and β2-glycoprotein 1 (β2-GP1)
Authors: Borchman D , Harris E N , Pierangeli S S , Lamba O P
Published: July 2010

β2-GP1 is a serum protein which influences binding of anticardiolipin antibodies to cardiolipin, may influence induction of these antibodies in animals and may play a role in anticardiolipin-mediated thrombosis. Various investigators have proposed that when β2-GP1 binds cardiolipin, structural alterations occur in one or both molecules, resulting in exposure of new epitopes for anticardiolipin binding, but there has been no proof that such alterations occur. Utilizing Fourier transform infrared spectroscopy, this study analysed the structure of cardiolipin and β2-GP1 alone, then mixed with each other. For pure cardiolipin, analysis of the CH2 stretching, scissoring and carbonyl bands suggested this molecule assumes a hexagonal crystal lattice packing structure in both anhydrous and aqueous samples. Based on the second derivative analysis of the amide 1 band from the β2-GP1 protein backbone, as well as Fourier self-deconvolution and curve fit algorithms, β2-GP1 was calculated to contain 18% turns, 37%α-helix, and 45%β-sheet structure. β2-GP1 binding with cardiolipin results in a significant change in the conformation as well as geometry of the lipid and protein components. This is indicated by a broadening of the CH stretching band and a marked shift in intensity of the carbonyl band of cardiolipin, indicating less hydrogen bonding. There was a decrease in β-sheet structure of β2-GP1 from 46% to 23% and appearance of 26% to 28% random structure. These findings indicate that mixing β2-GP1 with cardiolipin results in profound changes in both molecules which might explain the effect of β2-GP1 on anticardiolipin binding activity.
Differential alterations in plasma colony-stimulating factor concentrations in meningococcaemia
Authors: Waring P M , Presneill J , Maher D W , Layton J E , Cebon J , Waring L J , Metcalf D
Published: July 2010

To determine whether circulating levels of any of the cotony-stimulating factors (CSF) might contribute to the host response in severe sepsis, plasma concentrations of granulocyte CSF (GCSF), granulocyte-macrophage CSF (GM-CSF), and macrophage CSF (M-CSF), were measured by immunoassays in 20 subjects with meningococcaemia, a bloodstream infection caused by Neisseria meningitidis, that has proven to be a valuable model to study the responses of other inflammatory mediators during sepsis and septic shock in humans. Plasma G-CSF concentrations were transiently elevated in most subjects during the early phase of meningococcaemia, and were higher in subjects with septic shock (mean ± s.d. = 165± 142 ng/ml, n = 9) compared with those who remained normotensive (mean ± s.d.=7±2ng/ml, ng/ml, n=10) (p<005). Peak plasma G-CSF concentrations >10ng/ml were associated with the development of septic shock (P<0.01), disseminated intravascular coagulation (p<0.01). fulminant infection (P<0.05), and a fatal outcome (p<0.01). Plasma GM-CSF concentrations > 1 ng/ml were briefly present in subjects with life-threatening septic shock (1–15 ng/ml, n = 5). and were strongly associated with fulminant meningococcaemia (P <0.01). Plasma M-CSF concentrations were marginally elevated in all subjects, but were not associated with complications related to or arising from sepsis-induced organ injury. This study demonstrates that plasma levels of G-CSF, GM-CSF and M-CSF show very different responses during meningococcaemia, changes which presumably reflect the different roles played by these mediators in sepsis and, potentially, in septic shock.
Anti-malaria antibody-producing B cell frequencies in adults after aPlasmodium falciparumoutbreak in Madagascar
Authors: Migot F , Chougnet C , Henzel D , Dubois B , Jambou R , Fievet N , Deloron P
Published: July 2010

The central highlands of Madagascar offer a unique opportunity to explore the malaria immune memory, as the last murderous epidemic in the study area occurred 8 years ago. Quantification of the circulating memory B lymphocytes reacting to Plasmodium falciparum was assessed among 14 Madagascans by using a limiting dilution assay, applied to the EL4 culture system, which leads to activation, proliferation and differentia lion into antibody-secreting cells (ASC) of most peripheral B cells. This system allowed us to observe, without any malaria-specific restimulalion, a geometric mean frequency of one anti-P. falciparum ASC among 2992 circulating B cells, except for one Madagascan who did not have any detectable ASC. A geometric mean frequency of one anti-P. falciparum ASC among 1403 was obtained for six malaria hyperimmune Cameroonians, but conversely, no anti-malaria ASC was detected in the blood of six malaria non-immune French control subjects. Anti-P. falciparum ASC frequencies and serum specific antibodies were strongly related. Our results indicate that anti-malaria ASC are still present in peripheral blood of Madagascan subjects, who have not been exposed to P. falciparum for several years. These responder B cells reflect the malaria B cell memory acquired during the last epidemic.
Genetic variation in neutrophil accumulation in mice is not mediated through immigrant regulatory cells
Authors: Marley S B , Hadley C L , Wakelin D
Published: July 2010

Genetic variation of induced peritoneal neutrophilia in mice was accompanied by parallel variation in macrophage responses. The timing of the macrophage responses in high responder (C57B1/10) mice indicated a potential role for these cells in mediating the enhanced neutrophil response. However, adoptive transfer of inflammatory macrophages did not induce neutrophilia. Analysis of peritoneal cytokine levels in high and low responder mice further indicated [hat IL-1, IL-3, GM-CSE, G-CSE and interferon-gamma (IFN-γ) were not involved in mediating the genetic variation observed. Exogenous tumour necrosis factor-alpha (TNF-α) was effective in inducing the high responder phenotype, despite the absence of detectable TNF-α in either peritoneal fluid or serum. A role for genetically determined differential expression of endothelial adhesion molecules in high and low responders is suggested.
Impaired dendritic cell functions because of depletion of natural killer cells disrupt antigen-specific immune responses in mice: restoration of adaptive immunity in natural killer-depleted mice by antigen-pulsed dendritic cell
Authors: Yoshida O , Akbar F , Miyake T , Abe M , Matsuura B , Hiasa Y , Onji M
Published: July 2010

The primary aim of this study was to evaluate the role of natural killer (NK) cells on antigen-specific adaptive immune responses. After analysing the mechanism of impaired adaptive immune responses of NK-depleted mice, an immune interventional approach was developed to restore adaptive immunity in NK-depleted mice. NK cells were depleted from mice by administration of anti-asialo GM1 antibody (100 μl/mouse), twice, at an interval of 48 h. Hepatitis B surface antigen (HBsAg) was administered intraperitoneally to normal C57BL/6 mice (control mice) and NK-depleted mice. The levels of antibody to HBsAg (anti-HBs) in the sera and HBsAg-specific lymphocytes in the spleen were assessed. The functions of T lymphocytes, B lymphocytes and dendritic cells (DCs) were evaluated in vitro. HBsAg-pulsed DCs were prepared by culturing spleen DCs with HBsAg for 48 h and administered once to NK-depleted mice. The levels of anti-HBs in the sera and HBsAg-specific lymphocytes were significantly lower in NK-depleted mice compared with control mice (P < 0·05). The functions of T and B lymphocytes were similar between control mice and NK-depleted mice. However, the functions of spleen DC and liver DC were significantly lower in NK-depleted mice compared with control mice (P < 0·05). Administration of HBsAg-pulsed DCs, but not HBsAg, induced HBsAg-specific humoral and cellular immune responses in NK-depleted mice. Our study suggests that cross-talk between NK cells and DCs regulates the magnitude of adaptive immunity. In addition, antigen-pulsed immunogenic DCs represent potent immune modulator even if subjects with diminished innate immunity.
Two-dimensional gel electrophoresis analysis in simultaneous influenza pneumonia and bacterial infection in mice
Authors: Kosai K , Seki M , Yanagihara K , Nakamura S , Kurihara S , Imamura Y , Izumikawa K , Kakeya H , Yamamoto Y , Tashiro T , Kohno S
Published: July 2010

Severe pneumonia is found in simultaneous influenza pneumonia and bacterial infection, and suggests a relationship with immunological mechanisms. Here, we performed two-dimensional gel electrophoresis to detect immunological molecules related to the fulminant pneumonia caused by influenza virus and Streptococcus pneumoniae co-infection in mice. We found two spots that were expressed strongly in co-infected mouse lungs, compared with S. pneumoniae or influenza virus singly infected mouse lungs. The spots were analysed by mass spectrometry, and identified as alpha-1 anti-trypsin (A1AT), known as an anti-protease for neutrophil-derived proteolytic enzymes, and creatine kinase, which reflects a greater degree of lung damage and cell death. A1AT expression was increased significantly, and proteolytic enzymes from neutrophils, such as neutrophil elastase, myeloperoxidase and lysozyme, were also secreted abundantly in influenza virus and S. pneumoniae co-infected lungs compared with S. pneumoniae or influenza virus singly infected lungs. These data suggest that A1AT may play a central role as a molecule with broad anti-inflammatory properties, and regulation of the neutrophil-mediated severe lung inflammation is important in the pathogenesis of co-infection with influenza virus and bacteria.
Up-regulated macrophage migration inhibitory factor protects apoptosis of dermal fibroblasts in patients with systemic sclerosis
Authors: Kim J-Y , Kwok S-K , Hur K-H , Kim H-J , Kim N S , Yoo S-A , Kim W-U , Cho C-S
Published: July 2010

Macrophage migration inhibitory factor (MIF) is a proinflammatory cytokine that has been demonstrated to regulate the apoptosis of several cell types. Dysregulated apoptosis of fibroblasts has been implicated in a variety of fibrotic diseases, including systemic sclerosis (SSc). In this study, we investigated the role of MIF in the apoptosis of dermal fibroblasts. The concentrations of MIF were measured in sera and in culture supernatants of peripheral blood mononuclear cells (PBMCs) and dermal fibroblasts by enzyme-linked immunosorbent assay. The degree of apoptosis was determined by colorimetric assay, and signalling pathways were examined by Western blot. The results showed that serum levels of MIF were significantly higher in patients with SSc (n = 47) than in healthy controls (n = 56). Stimulation of PBMCs by anti-CD3 and anti-CD28 increased the production of MIF by fourfold over the constitutive levels. SSc dermal fibroblasts produced higher amounts of MIF than normal dermal fibroblasts. When treated with sodium nitroprusside (SNP), SSc dermal fibroblasts showed a lower degree of apoptosis compared with normal dermal fibroblasts. Exogenous MIF (1–100 ng/ml) inhibited SNP-induced apoptosis of dermal fibroblasts dose-dependently. Both extracellular regulated kinase (ERK) inhibitor (PD98059) and protein kinase B (Akt) inhibitor (LY294002) almost completely blocked the inhibitory effect of MIF on apoptosis. Furthermore, MIF increased the expression of Bcl-2, phospho-ERK and phospho-Akt activity in dermal fibroblasts. Taken together, our data suggest that MIF released by activated T cells and dermal fibroblasts decreases the apoptosis of dermal fibroblasts through activation of ERK, Akt and Bcl-2 signalling pathways, which might be associated with excessive fibrosis in SSc.
Autopsy approach to stroke
Authors: Love Seth
Published: July 2010

Love S (2011) Histopathology58, 333ndash;351 Autopsy approach to stroke

Stroke is a major cause of morbidity and mortality but the brain and other relevant tissues are often examined only cursorily when stroke patients come to autopsy. The pathological findings and clinical implications vary according to the type of stroke and its location and cause. Large ischaemic strokes are usually associated with atherosclerosis of extracranial or major intracranial arteries but can be caused by dissection. Most small cerebral infarcts are caused by arteriosclerosis or, in the elderly, cerebral amyloid angiopathy (CAA). However, vasculitides and coagulopathies can cause a range of different patterns of ischaemic (and, occasionally, haemorrhagic) stroke. Global brain ischaemia, caused by severe hypotension or raised intracranial pressure, produces damage that is accentuated in certain regions and neuronal populations and may be confused with hypoglycaemic injury. The main cause of subarachnoid haemorrhage is a ruptured berry aneurysm but CAA, arteriovenous malformations and infective aneurysms are occasionally responsible. These can also cause parenchymal brain haemorrhage, although this most often complicates hypertensive small vessel disease. Sometimes the haemorrhage arises from a neoplasm. Performing an adequate autopsy in stroke requires proper preparation, awareness of the likely pathological processes, familiarity with intracranial vascular anatomy, careful gross examination and dissection, and appropriate use of histology.
PhoB regulates both environmental and virulence gene expression in Vibrio cholerae
Authors: Pratt Jason T. , Ismail Ayman M. , Camilli Andrew
Published: July 2010

Vibrio cholerae is a facultative pathogen that thrives in two nutritionally disparate environments, aquatic and human small intestine. Phosphate (Pi) is an essential nutrient that is limited in aquatic ecosystems and of unknown availability in the small intestine. Here, we show that the Pi (Pho) regulon, which is controlled by the Pi‐specific transporter (Pst) and two‐component system PhoBR, is required for V. cholerae survival in both environments, though for differing reasons. While induction of Pi acquisition systems including Pst is critical for survival in the aquatic environment, regulation of virulence genes by PhoB and not Pi transport per se is required for colonization of the small intestine. We show that PhoB regulates virulence genes by directly controlling expression of a key upstream transcriptional regulator, tcpPH. Thus, the Pho regulon includes virulence genes and represents a diverse gene set essential to pathogenic V. cholerae throughout its life cycle.
Solasodine glycoalkaloids: a novel topical therapy for basal cell carcinoma. A double‐blind, randomized, placebo‐controlled, parallel group, multicenter study
Authors: Punjabi Sangeeta , Cook L. J. , Kersey P. , Marks R. , Cerio R.
Published: July 2010

Objective  To assess the safety and efficacy of a 0.005% mixture of solasodine glycosides (Zycure) in the treatment of basal cell carcinoma.

Design  Double‐blind, randomized, and vehicle‐controlled, parallel group study.

Setting  Ten centers in the United Kingdom.

Participants  Male, n = 50; female, n = 44; age range, 32–95 years (Table 1).

Demographics

Characteristic
Treatment group

Zycure n = 62
Vehicle n = 32

Age (in years): mean
68.9
70.0

Sex: male
33
17

Sex: female
29
15

Race: Caucasian
62
32

Intervention  Ninety‐four patients were randomized on a 2 : 1 ratio (n = 62, Zycure; n = 32, vehicle). Histologically confirmed lesions were treated double blinded, twice daily under occlusion with Zycure or vehicle for 8 weeks. Patients were reviewed fortnightly for adverse effects and overall response. Successfully treated patients were followed up at six‐month intervals for a year.

Main outcome measures  The primary efficacy endpoint was histologically confirmed clearance of the basal cell carcinoma (2‐mm punch biopsy) at the end of 8‐week treatment.

Results  Efficacy (intention‐to‐treat population) at 8 weeks was 66% (41/62) in the Zycure group, compared to 25% (8/32) in the vehicle group (P < 0.001; Cochran–Mantel–Haenszel test). Ninety percent (37/41) of the Zycure group completed follow‐up at six‐month intervals for 1 year, of whom 78% (29/37) had no recurrence. There were no major treatment‐related adverse effects, although 10 patients in Zycure group did not complete the treatment protocol for various reasons.

Conclusion  We conclude that the solasodine glycoside cream Zycure is a safe therapy for basal cell carcinoma, with a cure rate of 66% at 8 weeks and 78% at 1 year follow‐up.
Progress in Rhesus Histocompatibility Typing Resulting from the Second International Nonhuman Primate Histocompatibility Workshop (1973)
Authors: NEEFE J. R. , BALNER H. , BARNES A. D. , FORD C. , ROOENTINE G. N. , VAN VREESWIJK W. , WARD F. E.
Published: July 2010

The Second International Nonhuman Primate Histocompatibility Workshop permitted comparison of rhesus monkey alloantisera developd in various laboratories on a single common panel of related and unrelated monkeys. Analysis of the data permits the conclusion that at least nine specificities are recognized by more than one laboratory, including six at the first locus and three at the second locus.