bluuebunny/crossref_metadata_2025 huggingface dataset:
Antibody penetration of viable human cells. I. Increased penetration of human lymphocytes by anti-RNP IgG
Authors: MA J , CHAPMAN G V , CHEN S-L , MELICK G , PENNY R , BREIT S N
Published: July 2010

Antibody penetration of viable cells and interaction with intracelluiar antigens may have major consequences for immunopaihological processes in connective tissue diseases. We have reported previously that antibody can penetrate viable human lymphocytes. To assess further the role of antinuclear antibodies in this process, peripheral blood lymphocytes (PBMC) were incubated with FITC-conjugated IgG fractions from sera containing anti-RNP (anti-RNP IgG), Ro(SS-A), La(SS-B) and dsDNA antibodies and control sera for 24 h. Using crystal violet to quench cell surface staining, intracellular fluorescence of viable lymphocytes was quantified on the flow cytomcter. It was noted that anti-RNP IgG entered 46.4±7.2% of lymphocytes which was significantly higher than anti-Ro(SS-A) (29.9±4.1%, P < 0.05), La(SS-B) (22.0±7.5%, P < 0.01) IgG and control IgG (28.8±2.1%, P < 0.05) and not statistically different from anti-dsDNA IgG (32.6±14.3%). Inhibition experiments showed that the increased number of cells penetrated by anti-RNP IgG was a specific process. Time-course studies showed that anti-RNP IgG entry into cells was different from pooled control IgG. With anti-RNP IgG, positive-staining lymphocytes gradually increased in number from 12 to 24 h incubation, whilst with pooled control IgG, the peak was reached within 5 min. Dual staining experiments suggested that whereas both anli-RNP IgG and pooled control IgG entered Band NK. cells, anti-RNP IgG also entered T cells. Using IgG F(ab')2 and Fc fragments from either anti-RNP IgG or pooled control IgG to compete with their FITC-conjugated counterparts indicated that the entry of anti-RNP IgG into viable cells appeared to involve both F(ab')2 and Fc fragments, and pooled control IgG depended exclusively on the Fc portion of IgG. Further investigation by incubating anti-RNP IgG with 35S-methionine-labelled monocyte-depleted PBMC (MD-PBMC) suggested that anti-RNP IgG might react with the corresponding antigens either on the cell surface or within the cytoplasm.
Detection of endogenous cytokines in sera or in lymph nodes obtained from patients with sarcoidosis
Authors: ASANO M , MINAGAWA T , OHMICHI M , HIRAGA Y
Published: July 2010

To investigate the possible role of endogenous cytokines in the immunopathogenesis of sarcoidosis, a study of cytokines in lymph nodes obtained from patients with active pulmonary sarcoidosis was carried out using immunoperoxidase staining and radioimmunoassays (RIA). Cells stained for inlerfcron-y (IFN-γ) and tumour necrosis factor-α (TNF-α), which appeared to be CD3+ cells and CD14+ cells, respectively, were seen scattered around granulomas. In contrast, cells positive for intcrlcukin-1β (IL-1β) were located in CD11c+ cells within granulomas. Lymph nodes of patients with sarcoidosis contained significantly higher amounts of IFN-y, TNF-a and IL-1β than control lymph nodes. The levels of IFN-γ and TNF-α in extracts of lymph nodes were correlated with the histological status of the granulomas. IFN-γ was delected in all stages, while the highest level of TNF-α was obtained from lymph nodes containing many active granulomas. The level of serum IFN-γ was always lower than in lymph node extract and correlated significantly with either serum angiotensin-converting enzyme or serum lysozyme. Patients with positive gallium-67 uptake or bilateral hilar lymphadenopathy had high levels of either serum IFN-γ or lysozyme. These results suggest that quantitative analyses of IFN-γ and TNF-α in sera and lymph nodes may serve to elucidate the pathophysiology of sarcoidosis.
Autoantibodies to DNA topoisomerase II in juvenile rheumatoid arthritis
Authors: ZUKLYS K L , SZER I S , SZER W
Published: July 2010

Sera from 58 children with juvenile rheumatoid arthritis were examined for the presence of antibodies to DNA topoisomerase II. Eight sera were reactive in immunoblotting with purified human topoisomerase II and a protein encoded by a cloned cDNA expressed in Escherichia coli which represents the carboxy-terminal domain of the human enzyme. In addition, the sera detect topoisomerase II in mitotic chromosomes and chromosome scaffoids. Five of the sera bind to the native enzyme in solution and deplete such solutions of the active enzyme. All eight sera also contain antibodies to nuclear antigens other than topoisomerase II.
Differential staining of human αβ and γδ T cells by the fluorescein conjugate of an anti-CD3 monoclonal antibody
Authors: MULLERSMAN J E , WHITE G , TUNG K S K
Published: July 2010

The enumeration of total T cells, an important function of the clinical immunology laboratory, utilizes antibodies to CD3, the macrornolecular complex associated with the antigen-specific receptors of T cells. We compared the ability of some commonly employed commercial anti-CD3 reagents to stain human peripheral blood lymphocytes. Surprisingly, the fluorescein isothiocyanate (FTTC) conjugate of Coulter clone T3 (FITC-T3) stained most T cells brightly, but selectively stained γδ T cells very dimly or not at all. In contrast, the other anli-CD3 reagents studied (FITC-Leu 4, PE-T3, PE-Leu 4, and indirectly labelled T3 and Leu 4) stained all T cells equivalently. Dual-colour flow cylometric analysis wilh FITC -T3 and PE-Leu 4 readily demonstrated a F1TC-T3 /PE-Leu 4+ population of T cells. This unique population stained dimly or not at all with a combination of anti-CD4 and anti-CD8 monoclonal antibodies and positively with the pan-γδ T cell antibody TCR(5I. Moreover, an excellent correlation was found between the number of FITC-T3 /PE-Leu 4+ cells and the number of TCRδ1+ cells in 32 normal individuals. Thus, the FITC-T3- /PE-Leu 4+ phenotype accurately marks all γδ T ceils. In contrast to FITC T3. both PE-conjugated and unconjugated T3 stained γδ T cells brightly. Therefore, T3 binds to an epitope present on all T cells, but fluoresceinylalion specifically attenuates this antibody’s ability to bind to γδ T cells. These findings indicate that the use of FITC-T3 can result in a significant and variable underestimation of peripheral blood T cell number and demonstrate further that the CD3 complexes of human αβ and γδ T cells are significantly different.
Increased levels of soluble low-affinity Feγ receptors (IgG-binding factors) in the sera of tumour-bearing mice
Authors: LYNCH A , TARTOUR E , TEILLAUD J L , ASSELAIN B , FRIDMAN W H , SAUTÉS C
Published: July 2010

Soluble forms of low affinity Feγ receptors (FcyR), also called IgG-binding factors (IgG-BF), have been shown to play a regulatory role in immune responses. By using an immunodot assay with the anti-mouse FcyR MoAb, 2.4G2, the levels of IgG-BF have been measured in the sera of mice bearing syngeneic tumours of lymphoid or non-lymphoid origin or in mice injected with high doses of murine IgG. These sera contained large amounts of IgG-BF as compared with controls. In the case of mice bearing IgG2a- or IgG2b-secreting hybridomas or lymphomas, serum IgG-BF increased progressively with tumour size and serum monoclonal IgG concentration, reaching 4–12 times the normal levels. A less than three-fold increase was found in mice bearing an IgG1-secreting hybridoma or tumours which do not secrete IgG (IgA-secreting hybridoma, non-immunoglobulin-secreting lymphoid tumours or melanoma) or in mice injected with 9 mg of monoclonal IgG2a. The enhancement of serum IgG-BF levels was independent of the expression of FcγR by the tumour cells, suggesting that the majority of IgG-BF secreted in response to tumours was produced by the host rather than by the tumour. The increased production of IgG-BF may participate in the control of tumour growth and in the modulation of the host immune responses in tumour-bearing animals.
Thymus lymphocytes in uraemic rats and the effect of thymosin fraction 5in vivo
Authors: IKEMOTO S , KAMIZURU M , HAYAHARA N , OKAMOTO S , WADA S , KISHlMOTO T , MAEKAWA M
Published: July 2010

Thymus lymphocyte subsets in uraemic rats were studied using monoclonal antibodies. Severe and moderate uraemia was induced in rats, and sham-operated and normal rats were used as the controls. As a result, the weight of the thymus decreased in uraemic rats. As for lymphocyte subsets, the frequency of W3/25+OX8+ decreased and those of W3/25−OX8−, W3/25+OX8− and W3/25−OX8+ relatively increased in uraemic rats. All these changes were more significant in severely uraemic than in moderately uraemic rats. When thymosin fraction S (TF5) was administered to severely uraemic rats, the weight of the thymus increased and the lymphocytes subsets normalized. These results suggest that uraemia may cause a maturational impairment of thymus lymphocytes by the depression of thymic hormone secretion.
Study of CYP2D6 gene in children with autoimmune hepatitis and P450 IID6 autoantibodies
Authors: YAMAMOTO A M , MURA C , MORALES M G , BERNARD O , KRISHNAMOORTHY R , ALVAREZ P
Published: July 2010

Cytochrome P450 IID6 is an autoantigen recognized by the sera of children affected with a subtype of autoimmune hepatitis. It was hypothesized that a mutation in the CYP2D6 gene could explain the autoimmune response in these patients. To examine this question, genomic DNA from peripheral lymphocytes (n= 9) and liver (n=l) of 10 patients with anti-LKM-l antibody was analysed by Southern blot for genetic association studies between a particular CYP2D6 haplotypc and autoimmune hepatitis. In addition, a region of CYP2D6. from the same genomic DNA. was amplified by polymcrasc chain reaction (PCR) and digested by BstNI, in a search for the most prevalent 29B mutation, described in subjects who do not express the P45O IID6. Total RNA and proteins, prepared from the liver of an anti-LKM-l+ patient, were analysed by Northern and Western (immunoblot) blots respectively. Our results do not reveal any major structural change in the DNA of this patient at the CYP2D6 locus that could explain their autoimmune response. Corroborating this observation, no changes were noted either in P450 IID6 mRNA size or in the corresponding protein. However, these data do not exclude the possibility of subtle changes in the protein due to point mutations in critical regions that might trigger an autoimmune response.
Antigenic specificity and subset analysis of T cells isolated from the bronchoalveolar lavage and pleural effusion of patients with lung disease
Authors: FAITH A , SCHELLENBERG D M , REES A D M , MITCHELL D M
Published: July 2010

Cellular infiltraicsofbronchoalvcolarlavage(BAL)andpleuralcffusion from patients with tuberculosis (TB) and lung cancer were characterized for the presence of different T cell subsets by phenotypic analysis. The specificity of ihe T cells for mycobacierial antigens was then compared for the two disease compartments. The composition ofT cell subsets within the BAL. in contrast to pleural effusion cells (PEC), revealedevidenceofsequestrationofCD8’ cells. BALTcellswerefoundtobeapredominantly CD29’ DR’ memory population ofactivatcd cells. Although polyclonal populations of BAL T cells proliferated poorly to Mycohacterium tuberculosis antigens, mycobacierial antigen-reactive monoclonal T cell populations could be derived from the alveolar compartment. Two clones were shown to recognize the 65-kD heat shock protein of mycobactcria. and one of these clones recognized a conserved sequence of the molecule. Several BAL-derivcd clones, responding to a mycobacterial soluble extract, did not. however, recognize purified mycobacterial antigens, previously identified as highly stimulatory for PEC-derivcd T cells. T cell clones, derived from PEC of two TB patients, responded to the 38-kD and 71-kD. as well as the 65-kD mycobacterial antigens. Examination of the activation requirements of BAL-derivcd T cell clones, specific for mycobacterial antigens, revealed that exogenous IL-2 was necessary for the T cells to sustain proliferation. This was in contrast to the mycobacterial antigen-reactive T cells cloned from PEC. These results suggest that T cell populations with distinct antigen specificities and activation requirements arc present in BAL and PEC.
HBsAg in urine: a new approach for the detection of urinary antigens
Authors: BADUR S , GRANGEOT-KEROS L , PILLOT J
Published: July 2010

In order to define the optimal conditions for detection of microbial antigens in urine, urinary HBsAg excreted during hepatitis B was chosen as a model. Using commercial kits, which mainly involve anti-discontinuous epitopes, we found urinary HBsAg in only 50% of patients with HBsAg in their sera. In contrast, with an inhibition method involving a monoclonal antibody recognizing a continuous epitope, urinary HBsAg was found in 100% of these patients. Structural analysis of HBsAg showed that urinary HBsAg is denatured; it can escape detection by commercial kits well fitted for detection of native serum HBsAg. General implications for the revelation of urinary microbial antigens are discussed.
Interaction and intracellular killing ofCandida albicansblastospores by human polymorphonuclear leucocytes, monocytes and monocyte-derived macrophages in aerobic and anaerobic conditions
Authors: THOMPSON H L , WILTON J M A
Published: July 2010

Polymorphonuclear leucocytes (PMN), monocytes and monocyte-derived macrophages were capable of interacting with opsonized C. albicans in both aerobic and anaerobic conditions. Superoxide anion release by these cells was inhibited in anaerobic conditions while lysozyme release and phagocytosis were equally efficient in both aerobic and anaerobic conditions. All cell types tested were capable of intracellular killing of C. albicans and this appeared to be maximum at 6 h for monocytes and macrophages and 24 h For PMN. Monocytes killed the lowest number of organisms, 1·106, and the killing was similar for aerobic and anaerobic conditions. In contrast, PMN and macrophages demonstrated greater killing of C. albicans in aerobic conditions compared with anaerobic conditions: PMN killed 1.9 ± 106 organisms and macrophages 3 ± 106 when incubated anaerobically. Inhibitors of oxygen metabolism decreased intracellular killing of C. albicans by macrophages and PMN in aerobic but not anaerobic conditions. The oxygen reaction products involved in the killing of C. albicans appeared to be different however: macrophage killing was decreased by superoxide anion and hydrogen peroxide inhibitors. PMN killing was decreased by superoxide anion, hydrogen peroxide, hypochlorous acid and hydroxyl radical inhibitors. The present study shows that although monocytes, macrophages and PMN function similarly in their interaction with C. albicans, they appear to use different oxygen reactive products for the intracellular killing of C. albicans.
Cell phenotypes in the efferent lymph of sheep persistently infected with Border disease virus
Authors: ENTRICAN G , HOPKINS J , MACLEAN M , MCCONNELL I , NETTLETON P F
Published: July 2010

The prefemoral efferent lymphatics of sheep persistently infected (PI) with Border disease virus (BDV) were cannulated in order to study the effects of the virus on cells of the immune system. Efferent lymphocytes recovered from PI sheep were phenotyped using a panel of monoclonal antibodies (MoAb) specific for ovine cell-surface markers and compared to lymphocytes recovered from normal, healthy controls. PI sheep had an increased percentage of cells expressing the T cell-associated molecules CD5, CD4, CD8 and TI9, also an increase in cells expressing CD1 and a population of cells expressing low levels of the T19 molecule which was not present in control sheep. The lymphocytes were examined for the presence of BDV using virus-specific MoAb. On average 8.5% of the efferent lymphocytes from PI sheep carried virus antigen. BDV antigen was also found in the mononuclear cells and connective tissue of lymph nodes indicating widespread virus dissemination within the lymphoid system of PI sheep.
Peripheral catabolism of CR1 (the C3b receptor, CD35) on erythrocytes from healthy individuals and patients with systemic lupus erythematosus (SLE)
Authors: COHEN J H M , LUTZ H U , PENNAFORTE J L , BOUCHARD A , KAZATCHKINE M D
Published: July 2010

The present study investigated the rate of catabolism of CR1 (the C3b receptor, CD35) on erythrocytes (E) in vivo, in relationship with the expressed number of CR1/E, the CR1.1. Hin dIII quantitative CR1 polymorphism, and cell age. The relationship between the number of CR1/E and cell age was analysed by measuring G6PDH activity in E that had been sorted according to high or low expression of CR1 (CD35), by assessing the expression of CR1 (CD35)on E separated according to cell density, and by comparing the number of CR1 (CD35) antigenic sites on reticulocytes and on E. A physiological catabolism of CR1 (CD35) manifested by a reduction in the number of CR1 (CD35) antigenic sites/E with cell ageing was consistently observed in healthy individuals. The number of CR1/E decreased with ageing of E according to a complex pattern that associated an exponential decay and an offset. Calculated half-lives of CR1 (CD35) ranged between 11 and 32 days in healthy individuals. A more rapid loss of CR1 (CD35) with cell ageing occurred on cells from individuals expressing high numbers of CR1/E. In patients with systemic lupus erythematosus (SLE). half-lives of CR1 (CD35) on E were in the same range as those of healthy individuals with a similar quantitative CR1 genotype; the number of CR1 (CD35) on reticulocytes was reduced and linearly related to the number of CR1/E, independently of the patients' quantitative CR1 genotype. Transfusion experiments with E bearing high or low amounts of CR1/E indicated the lack of preferential removal of E bearing high numbers of CR1 (CD35) in patients with SLE. These results indicate that the rate of loss of CR1 (CD35) from E with cell ageing is directly related to the quantitative CR1 phenotype and suggest that enhanced peripheral catabolism is not the sole mechanism of the acquired loss of CR1 (CD35) on E in patients with SLE.
Abnormal urothelial HLA-DR expression in interstitial cystitis
Authors: CHRISTMAS T J , BOTTAZZO G F
Published: July 2010

Interstitial cystitis is a chronic inflammatory disorder of the urinary bladder that predominantly afflicts middle-age women. The end stage of the disease is ulceration of the urothelium, the so-called Hunner's ulcer. The aetiology of interstitial cystitis remains obscure. We have studied bladder biopsies from 22 cases of interstitial cystitis and control groups consisting of six cases of bacterial cystitis and eight healthy women. Indirect immunofluorescence was performed on the biopsies using murine MoAbs to human HLA class I molecules, and class II molecules, HLA-DP, HLA-DQ and HLA-DR. In interstitial cystitis, bacterial cystitis and normal controls most cells expressed HLA class I products. In six cases of interstitial cystitis and one case of bacterial cystitis there was evidence of HLA class I hyperexpression. In normal bladder and bacterial cystitis HLA class II expression was restricted to submucosal dendritic cells. Langerhans cells macrophages, vascular endothelial cells and activated lymphocytes. All but two cases of interstitial cystitis showed surface expression of HLA-DR (but not HLA-DP or DQ). In all cases of interstitial cystitis there was an increase in the numbers of macrophages, activated lymphocytes and vascular endothelial cells expressing HLA class II molecules within the submucosa. These findings provide further evidence for the importance of inappropriate HLA molecule expression in a disease suspected of having an autoimmune pathogenesis and where cellular autoimmune mechanisms play a decisive role in the destruction of the target cells - the bladder urothelium.
Suppression of efferent limb of testicular autoimmune response by a regulatory CD4+ T cell line in mice
Authors: ITOH M , MUKASA A , TOKUNAGA Y , HIRAMINE C , HOJO K
Published: July 2010

A murine T cell line (designated as C.Ts) as a mediator of suppression of experimental autoimmune orchitis (EAO) was established. The method of establishment of C.Ts cell line was preparing spleen cells from C3H/He mice hyperimmunized with testicular germ cells (TC) and the repeated selection of the lymphocytes in vitro by stimulation with mouse testicular antigens (mTA). The C.Ts cells were Thy 1.2+, surface immunoglobulin, CD3+, CD4+ and CD8−. The cells could suppress the induction of EAO when transferred into actively EAO-sensitized mice only at the pre-clinical stage of the disease (efferent limb of the autoimmune response). The transferred C.Ts cells significantly inhibited both cellular and humoral immune responses to TC in the recipients in an antigen-specific manner. The disease suppression by C.Ts cells was found to depend upon their cell number, and their suppressive activity was markedly augmented by in vitro stimulation with mTA.
The structure of two IgMs showing different activity from a patient with Waldenstrom's macroglobulinaemia
Authors: MÉNDEZ E , OSUNA C , SÁNCHEZ A , REVILLA Y , SORIANO F , MONTALBÁN C , SEGUÍ J , AVILA J
Published: July 2010

The two monoclonal IgMs (IgM1 and IgM2) were characterized from a patient with Waldenström's macroglobulinaemia that resulted in a gammapathy. Heavy and light chains were isolated from the IgM. The complete primary structure of the two light chains and the NH2-terminal region of the two heavy chain molecules were determined. The sequence data indicated that the heavy and light chains from both IgMs belong to the same (III and II) lambda subgroups. By testing their antibody activity it was found by immunofluorescence and immunoblotting that only IgM2 reacts with an intermediate filament protein.
Induction of lymphocyte proliferation by antigen-pulsed human neutrophils
Authors: PRIOR C , TOWNSEND P J , HUGHES D A , HASLAM P L
Published: July 2010

We have investigated whether purified antigen-pulsed human neutrophils can induce a proliferative response in purified resting blood lymphocytes. Neutrophils were pulsed with soluble tetanus toxoid (dose range 25–250 Lf/ml) and co-cultured with autologous lymphocytes that had been depleted of MHC class II expressing cells. The antigen-pulsed neutrophils induced an increase of lymphocyte proliferation which was dependent on the antigen dose and the neutrophil/lymphocyte ratios. Neutrophils were less potent than autologous monocytes in stimulating lymphocyte proliferation. Blocking with a monoclonal antibody to a common determinant of the human MHC class II complex failed to reduce the lymphoproliferative effects and allogenic antigen-pulsed neutrophils were also able to elicit lymphocyte proliferation similar to autologous neutrophils. We conclude that antigen-pulsed neutrophils are able to induce lymphocyte proliferation in a non-MHC-restricted fashion.
Vaccination and genetic experiments demonstrate that adjuvant-oil-induced arthritis and homologous type II collagen-induced arthritis in the same rat strain are different diseases
Authors: HOLMDAHL R , KVICK C
Published: July 2010

The DA rat is highly susceptible to induction of arthritis after immunization with homologous type II collagen (CII) emulsified in Freund's incomplete adjuvant (FIA), resulting in collagen-induced arthritis (CIA). The DA rat also develops arthritis after injection of FIA alone (oil-induced arthritis (OIA)). This finding allows a direct comparison of two different models for rheumatoid arthritis; one induced with a defined auto-immunogen and one with a pure adjuvant. Both CIA and OIA develop approximately 2 weeks after induction but OIA is a self-limited acute disease whereas CIA induced with homologous CII follows a chronic disease course. Immunization with CII leads to a strong autoantibody response to CII while injection of FIA leads to no or very limited anti-CII antibody response. The Lewis rat develops neither CIA nor OIA while FI(DA ± Lewis) rats develop CIA but not OIA. Olive oil or CII emulsified in olive oil does not induce arthritis in DA rats. Pretreatment with CII in olive oil vaccinates against CIA but not OIA whereas pretreatment with FIA vaccinates against OIA but not CIA. These findings demonstrate that inclusion of CII in the adjuvant leads to a disease distinct from OIA which is characterized by a CII autoimmune response and chronicity of the disease course.
Modulation of the cellular immune response during Plasmodium falciparum infections in sickle cell trait individuals
Authors: ABU-ZEID Y A , THEANDER T G , ABDULHADI N H , HVIID L , SAEED B O , JEPSEN S , JENSEN J B , BAYOUMI R A
Published: July 2010

Plasma and peripheral blood mononuclear cells (PBMC) were obtained from P. falciparum-infecled individuals with and without the sickle cell trait at diagnosis and 7 days after treatment. I lbAA and HbAS patients were compared for levels of plasma soluble IL-2 receptors (IL-2R) and the in vitro cellular reactivity to affinity-purified soluble P. falciparum antigens (SPAg). PPD and phytohae-magglutinin (PHA). At diagnosis. HbAS patients with clinical disease had lower plasma-soluble IL-2R levels and parasite counts than the corresponding HbAA patients, whereas HbAS and HbAA patients with asymptomatic infections had comparable soluble IL-2R levels and parasite counts. PBMC from HbAS patients had higher proliferation and IFN-γ production in response to SPAg than PBMC from HbAA patients. The difference in the lymphoproliferativc responses to SPAg between the two groups was evident in patients with asymptomatic infections. In all patients, the clinical severity, the soluble IL-2R levels and the parasite counts were directly related. The former two were inversely related to the proliferative responses to SPAg. After treatment, all the studied parameters were comparable in the two groups. The study indicates that during P. falciparum infection, HbAS compared with HbAA patients had lower in vivo cellular activation and higher in vitro cellular reactivity in response to soluble malaria antigens.
Expression of the 40 kD protein in DLD-1 colon cancer cells and the effect of cytokines
Authors: DAS K M , SQUILLANTE L , ROBERTSON F
Published: July 2010

We recently reported the presence of an organ-specific 40 kD colonic protein which acts as an autoantigcn(s) in patients with ulcerative colitis. Using a specific monoclonal antibody directed against 40 kD protein (7E12H12, IgM isotype), in conjunction with immunocytochemistry and flow cytometry. we examined the presence of the 40 kD protein on human colon cancer cells. DLD-1, and also characterized the ability of cytokines. IFN-y and tumour necrosis factor, to modulate the expression of this protein on these tumour cells. The presence of the 40 kD protein was localized to the plasma membrane; less was present within the cytoplasm. Following exposure to IFN-γ (10–1000 Lty ml). DLD-1 colon tumour cells showed a dose- and time-dependent increase in 7E12H12 antibody associated immunofluorescence. with the maximum 7H12H12 antibody binding observed with 100 U/ml IFN-γ at 48 h. In contrast, tumour necrosis factor did not alter the levels of anti-40 kD antibody binding over that of control cells. Since IFN-γ is also known to induce class II major histocompatibility antigens, we examined the possibility of cross-reactivity of HLA class II antigens and Mr40 kD epitope. Neither pre-incubation of DLD-1 colon tumour cells with anti-HLA class II antibodies followed by 7E12H12 nor co-incubation of both antibodies altered the amount of 7E12H12 antibody binding. Using a direct ELISA, a highly enriched preparation of M, 40 kD protein reactive to anti-40 kD antibody did not react with HLA class II antibodies. The present results suggest that 40 kD protein is present on DLD-1 human colon tumour cells and that although the 40 kD protein epitope expression is increased by the lymphocyte-derived cylokine, IFN-y. the epitope is separate and distinct from the class II HLA antigens. Further studies on the 40 KD protein may elucidate its autoantigenic role in the pathogencsis of inflammatory bowel disease.
Acute rejection in heart transplant patients is associated with the presence of committed donor-specific cytotoxic lymphocytes in the graft but not in the blood
Authors: VAESSEN L M B , BAAN C C , OUWEHAND A J , JUTTE N H P M , BALK A H M M , MOCHTAR B , CLAAS F H J , WEIMAR W
Published: July 2010

In vivo-activated, commuted donor-specific cytotoxic lymphocytes (cCTL) can be propagated and expanded from endomyocardial biopsies (EMB) in IL-2-enriched medium especially during an acute rejection episode. We report here our efforts to detect these cCTL by the same technique in peripheral blood at the moment of rejection and when no rejection was diagnosed. During or just before rejection, significantly less frequent (P < 0·01) donor reactive cCTL were found in PBL samples (two out of 20) than in the simultaneously taken EMB samples (13 out of 19). Donor B-LCL and/or third-party B-LCL were lysed by 15 PBL samples. Inhibition studies revealed that this lysis was due to LAK-like cytotoxicity. The results show that peripheral blood does not reflect intra-graft events, which is probably the reason for the irrcproduciblc results of diagnosis of rejection by monitoring immunological parameters in the peripheral blood.
Specificity of IgG subclass antibodies in different clinical manifestations of leprosy
Authors: DHANDAYUTHAPANI S , IZUMI S , ANANDAN D , BHATIA V N
Published: July 2010

We analysed specific IgG subclasses levels to Mycobacterium leprae sonicate extract (MSE), lipoarabinomannan B (LAM) and phenolic glycolipid I (PGL-I) in the sera of leprosy patients with different clinical manifestations. IgG2 was found to be the predominant antibody to MSE regardless of clinical manifestations, and IgG 1 response was mostly seen in lepromatous patients. IgG3 reacted only rarely but IgG4 reacted relatively more in certain clinical groups such as borderline lepromatous and lepromatous with erythema nodosum leprosum (ENL) reaction. Most of the IgG subclass responses to MSE could be accounted for reactivity with LAM, suggesting that LAM is the major immunogen involved in the pathogenesis of leprosy. In contrast to LAM. PGL-I antigen showed considerably lower reactivities for IgG subclasses. An association between IgG subclass responses and clinical manifestations of leprosy was also seen. Whereas borderline lepromatous patients were found to have significantly higher levels of IgG2 and lgG4 to MSE, lepromatous patients had elevated levels of IgG I and lower levels of IgG2. An interesting observation, however, was the significantly higher levels of IgG2 to LAM in the pure neuritic leprosy patients.
The profile of IgG and IgG subclasses of onchocerciasis patients
Authors: DAFA'ALLA T H , GHALIB H W , ABDELMAGEED A , WILLIAMS J F
Published: July 2010

In this study Onchocerca gutturosa was compared with O. volvulus in an ELISA test to detect Onehocerca- spefic IgG and IgG subclasses. The test was developed and standardized to detect Onehocerca-spefic IgG and IgG subclasses in sera of onchocerciasis patients and endemic controls. Onchocerca volvulus and O. gutturosa crude water-soluble antigens showed no significant difference in detecting onchocerca-specific IgG antibody (T = 1·88, P > 0·05). The levels of IgG subclasses varied greatly. IgG4 showed the highest detected mean level (0·84 ± 0·59) and the other three subclasses showed considerably lower mean levels (IgGI =0·27±0·16, IgG2 = 0·24 ± 0·17, IgG3 = 0·28 ± 0·12). The status and score of skin lesions were found to have significant effect on the IgG and IgG subclasses levels (all P < 0·001). lgG4 showed a positive correlation with the microfilarial (Mf) load (r= 0·21, P < 0·03). IgG3 levels have a significant negative correlation with the Mf load (r=-0·23, P <0·02), The biological significance of these IgG and IgG subclasses in onchocerciasis is discussed.
Single and combined effects of the vitamin D analogue KH1060 and cyclosporin A on mercuric-chloride-induced autoimmune disease in the BN rat
Authors: LILLEVANG S T , ROSENKVIST J , ANDERSEN C B , LARSEN S , KEMP E , KRISTENSEN T
Published: July 2010

Mercuric chloride induces in BN rats a self-limiting systemic autoimmune disease characterized by proliferation of autoreactive CD4+ T lymphocytes, polyclonal activation of B lymphocytes, and the development of an anti-glomerular basement membrane (GBM) nephritis with concomitant nephrotic range protcinuria. We have used this model of autoimmune disease to test the immunosuppressive ability of a novel vitamin D3 analogue KH 1060. This compound prevents autoimmune manifestations including proteinuria. scrum IgE, and serum anti-laminin antibodies in a dose-dependent manner, as does cyclosporin A (CyA). When dosages of KH 1060 capable of partial reduction of proteinuria without causing significant hypercalcaemia are combined with small dosages of CyA also capable of partial prevention of protcinuria, an additive effect is seen, leading to complete prevention of proteinuria and substantial reductions in serum IgE and anti-laminin levels. Possible mechanisms of action are discussed and it is suggested that KH1060 could prove useful as an immunosuppressive agent in the treatment of autoimmune diseases.
Effect of interferon-gamma priming on the activation of murine peritoneal macrophages to tumouricidal state by cisplatin, IL-1, and tumour necrosis factor (TNF): production of IL-1 and TNF
Authors: SODHI A , SINGH R K , SINGH S M
Published: July 2010

The effect of interferon-gamma (IFN-γ) priming of murine peritoneal macrophages on the activation to tumouricidal state by cisplatin, lipopolysaccharide (LPS)-IL-1 and TNF was investigated. Cisplatin-, LPS-, IL-1- or TNF-treated IFN-γ-primed macrophages showed significantly enhanced tumouricidal activity and binding to tumour cells, compared with unprimed treated or untreated macrophages. Macrophages treated with cisplatin, LPS, IL-1 and TNF produced released and membrane-associated IL-1 and TNF activity which was significantly enhanced after priming with IFN-γ. These observations suggest the use of IFN-γ along with these biological response modifiers in designing immunotherapeutic protocols for treatment of malignancy.
Evaluation of serum levels of tumour necrosis factor-alpha (TNF-α) and soluble IL-2 receptor (sIL-2R) and CD4, CD8 and natural killer (NK) populations during infrared pulsed laser device (IPLD) treatment
Authors: SANTANA-BLANK L A , CASTES M , ROJAS M E , VARGAS F , SCOTT-ALGARA D
Published: July 2010

The purpose of this study was to evaluate serum levels of TNF-α, sIL-2R and distribution of peripheral leucocyte subsets in patients with advanced neoplastic disease undergoing IPLD treatment. Fifteen cancer patients with evidence of persistent disease were further divided in two groups according to outcome at the end of the period of clinical evaluation: group 1 patients were still alive and group 2 patients had died. Our results show: (i) an increase in the initial level of TNF-α in both groups; (ii) a decrease in TNF-α levels during the follow up of group I patients; (iii) a significant increase in serum levels of sIL-2R in patients in group 2 compared with those in group 1; (iv) a progressive and constant increase in TNF-α levels in group 2; (v) a decrease in CD4+CD45RA+ subpopulation in both groups; (vi) an increase in CD25+ cells; (vii) an increase in CD4+, CD4+ CD45RA+ and CD25+ cells during the follow up of group 2 patients. The data generated here form the basis for further investigations on the use of IPLD as a single agent and in combination with other biological response modifiers in cancer patients.
Tumour necrosis factor enhances the asbestos-induced production of reactive oxygen metabolites by human polymorphonuclear leucocytes (PMN)
Authors: KLOCKARS M , SAVOLAINEN H
Published: July 2010

We studied the effect of recombinant tumour necrosis factor alpha (TNF-α) on the production of reactive oxygen metabolites (ROM) by human PMN exposed in vitro to chrysotile and crocidolite asbestos fibres, quartz dusts and opsonized zymosan. TNF caused a significant increase in ROM release by PMN, and significantly and dose-dependently amplified the ROM production induced by asbestos fibres. The amplification of ROM production by TNF can be of crucial importance in the process of lung inflammation and fibrogenesis in pneumoconioses.
Absence of circulating interferon in patients with inflammatory bowel disease. Suggestion against an autoimmune etiology
Authors: CAPOBIANCHI M R , FAIS S , DI PAOLO M C , AGOSTINI D , PAOLUZI P , PALLONE F , DIANZANI F
Published: July 2010

Whether inflammatory bowel diseases (IBD) can be classified as autoimmune disorders is not established. Since circulating acid-labile interferon-alpha (IFN-α) is believed to reflect autoimmune reactions, we tested sera from two groups of IBD patients for the presence of circulating IFN. No detectable IFN was found in 51 serum samples of IBD patients. Furthermore, in no serum sample of IBD patients were neutralizing anti-IFN antibodies found. In contrast, acid-labile IFN-αwas present in sera from 21/52 HIV-infected and from 6/14 systemic lupus erythematosus patients. These observations provide evidence that IBD differs from systemic autoimmune disorders, at least for the presence of circulating IFN.
Renal allograft rejection: induction and function of adhesion molecules on cultured epithelial cells
Authors: LIN Y , KIRBY J A , CLARK K , SHENTON B K , FORSYTHE J L R , PROUD G , TAYLOR R M R
Published: July 2010

The interaction of graft-infiltrating immune cells with donor parenchymal cells is an important early event in allograft rejection. This binding is stabilized by interaction of antigen-independent ‘adhesion’ molecules expressed on the two cell types. As the level of expression of these molecules can be altered during inflammation, a series of experiments was performed to examine the effects of the inflammatory cytokines interferon-gamma (IFN-γ) and tumour necrosis factor-alpha (TNF-α) on adhesion molecules expressed by cultured human renal tubular epithelial cells. These cells constitutively expressed ICAM-1 and LFA-3. Incubation with IFN-γ increased expression of ICAM-1 but had no significant effect on expression of LFA-3 (P>0·05). Incubation with TNF-γ increased expression of both ICAM-1 and LFA-3; IFN-γ synergized with TNF-α to further augment expression of these molecules. Peripheral blood lymphocytes (PBL) showed an enhanced binding to allogeneic renal epithelial cell monolayers which had been pretreated with IFN-γ or TNF-α. MoAbs specific for ICAM-1 or its ligand LFA-1 inhibited adhesion of PBL to either IFN-γ- or TNF-α pretreated renal cells. By contrast, antibodies specific for LFA-3 or its ligand CD2 only significantly blocked PBL adhesion to renal cells which had been pretreated with TNF-α. Combination of antibodies specific for multiple components of the adhesion systems produced greater inhibition of adhesion than was produced by any single MoAb. These results suggest that the inflammatory cytokines IFN-γ and TNF-α up-regulate expression of functional ICAM-1 and LFA-3 molecules which can augment the binding of potentially graft-damaging lymphoid cells to renal tubular epithelial cells.
Mesangial sclerotic change with persistent proteinuria in rats after two consecutive injections of monoclonal antibody 1–22–3
Authors: KAWACHI H , IWANAGA T , TOYABE S , OITE T , SHIMIZU F
Published: July 2010

Irreversible mesangial changes with persistenl proteinuria were induced in rats given two consecutive injections 2 weeks apart of a MoAb 1–22–3 to rat mesangial cell. The characteristics of the resulting lesions were investigated and compared with those of the reversible change induced by a single injection. At 24 h after the second injection, mesangiolytic changes similar to those after a single injection were evident. The accumulation of macrophagc-like cells in glomeruli observed at I week after the first injection was not evident during the experimental period after the second injection. Hypercellularity with the characteristics of intrinsic mesangial cell and increased mesangial matrix were already present I week after the second injection. And mesangial sclerotic change progressed up to 6 months. Deposition of collagen type I and type III and accumulation of collagen fibril at the ullraslructural level were evident in rats 6 months after the second injection. Proteinuria started immediately and continued for more than 6 months after the second injection. The mesangial sclerotic change with persistent proteinuria described here is considered to be a better model for investigating the mechanism of chronic progression of human mesangial proliferative glomerulo-nephritis.
CD4-independent binding of HIV-1 to the B lymphocyte receptor CR2 (CD21) in the presence of complement and antibody
Authors: MONTEFIORI D C , ZHOU J , SHAFF D I
Published: July 2010

Complement and antibody contribute to infection-enhancement and possible expanded cellular tropism of HIV-1 in vitro through a process requiring complement receptors. Until now, however, the ability of HIV-1 to bind complement receptors has not been documented or characterized. We investigated whether antibody and complement permitted HIV-1 to bind to the B lymphocyte receptor, CR2 (CD21), in an effort to learn more about infection-enhancement, and also because CR2 can mediate B cell proliferation and antigen localization in lymphoid organs in other systems. HIV-1 incubated with antibody and fresh human serum as a source of complement bound approximately 10-fold greater to cells expressing CR2 than to HIV-1-permissive cells lacking this receptor. A similar effect was observed using cells which expressed CR2 but no CD4. This binding was minimal in heat-inactivated and C3-deficient sera, and was significantly reduced by the anti-CR2 MoAb, OKB7, but not by the anti-CD4 MoAb, OKT4a. Thus, complement and antibody acted in concert to facilitate the binding of HIV-1 to CR2 independently of CD4. CD4-independent binding of HIV-1 to CR2 was not sufficient to produce infection in Raji-3 cells. Titres of antibodies mediating CR2 binding correlated with antibody titres as measured by immunofluorescenee (P < 0.01) and infection-enhancement (P < 0.05) but were discordant with titres of neutralizing antibodies, a result consistent with the utilization of CR2 for enhanced infection of cells. The ability of complement and antibody to facilitate the binding of HIV-1 to CR2 in the absence of CD4 provides new insights into mechanisms of HIV-1-induced immunopathogenesis and infection-enhancement.
Cystic ovaries in women affected with hereditary angioedema
Authors: PERRICONE R , PASETTO N , DE CAROLIS C , VAQUERO E , NOCCIOLI G , PANERAI A E , FONTANA L
Published: July 2010

Polycystic ovary (PCO) syndrome is biochemically characterized by abnormal gonadotropin secretion and polycystic ovaries associated with increase in size and functional activity of stromal tissue; multifollicular ovaries (MFO) are defined by the presence of multiple cysts with no increase in stromal tissue. A central (hypothalamic-pituitary) abnormality, including high plasma /?-endorphin (BE) concentrations without simultaneous elevation of ACTH, was reported for subjects with PCO syndrome. Since we have found the presence of high plasma BE concentrations in hereditary angioedema (H ANE) during attacks as well as during symptom-free periods, we studied, by means of pelvic ultrasound scanning employed to determine the prevalence of PCO and of MFO, 13 women of reproductive age affected with HAN E who were not on oral contraceptives. We have found PCO in 5/ 13 (38.4%) and MFO in 7/13 (53.8%) HANE patients. Nine patients had oligomenorrhoea (five with PCO, three with MFO, one with normal ovaries), five (three with PCO, two with MFO) were hirsute and only one (with MFO) had weight loss. No patient was obese. Mean plasma LH, testosterone, prolactin, cortisol and ACTH concentrations were normal, while FSH was significantly reduced and LH/FSH ratio increased. BE concentrations were significantly high in all the patients studied. Our results clearly demonstrate that women with HANE frequently have cystic ovaries (polycystic or multifollicular) in the presence of high BE concentrations.
An altered repertoire of T cell receptor V gene expression by rheumatoid synovial fluid T lymphocytes
Authors: LUNARDI C , MARGUERIE C , SO A K
Published: July 2010

The pattern of T cell receptor V gene expression by lymphocytes from rheumatoid synovial fluid and paired peripheral blood samples was compared using a polymerasc chain reaction (PCR)-based assay. Eight rheumatoid arthritis (RA) patients who had varying durations of disease (from 2 to 20 years) were studied. In all patients there was evidence of a different pattern of V gene expression between the two compartments. Significantly increased expression of at least one Vα or Vβ gene family by synovial fluid T cells was observed in all the patients studied. Three different Vα (Vα 10, 15 and 18) and three Vβ (Vβ 4, 5 and 13) families were commonly elevated. Sequencing of synovial Vβ transcripts demonstrated that the basis of increased expression of selected V gene families in the synovial fluid was due to the presence of dominant clonotypes within those families, which constituted up to 53% of the sequences isolated from one particular synovial V gene family. There were considerable differences in the NDJ sequences found in synovial and peripheral blood T cell receptor (TCR) transcripts of the same Vα gene family. These data suggest that the TCR repertoire in the two compartments differs, and that antigen-driven expansion of particular synovial T cell populations is a component of rheumatoid synovitis, and is present in all stages of the disease.
Glomerular immune deposits in murine lupus models may contain histones
Authors: SCHMIEDEKE T , STOECKL F , MULLER S , SUGISAKI Y , BATSFORD S , WOITAS R , VOGT A
Published: July 2010

Two types of lupus mice, NZB/NZW F1 female hybrids and mice with graft-rersus-host disease (GVHD). were studied. Histones H3 and H2A were detected by immunofluorcscenee in glomeruli of 22/22 proteinuric GVHD and 8/12 proteinuric NZB/W F1 female mice; in non-proteinuric animals. 3/5 GVHD and 2/27 NZB/W F1 female were positive. Using antibodies to histone peptides it was shown that mainly the N-terminal regions of histones H3 and H2A were exposed in glomerular deposits. Western blot analysis revealed antibodies lo histone subfractions in sera of 33/34 lupus mice that developed proteinuria. This study provides evidence that histones are involved in the pathogenesis of lupus nephritis.
Immunization of mice with mycobacterial culture filtrate proteins
Authors: HUBBARD R D , FLORY C M , COLLINS F M
Published: July 2010

Culture filtrate proteins were obtained from Mycobacterium tuberculosis cultures after 7 days growth in Proskauer and Beck medium. The protein yield increased substantially to peak about the time the number of viable organisms reached its maximum level (day 8). Examination of the protein concentrate by SDS PAGE revealed the presence of at least 12 separate protein bands varying from 10 to 90 kD. Mice were injected subcutaneously with 20μg of M. tuberculosis culture filtrate (MTCF) protein suspended in saline or Freund's complete or incomplete adjuvant. The vaccinated mice were subjected to an aerogenic challenge with 103 colony-forming unit (CPU) M. tuberculosis Erdman and a significant reduction in the number of viable organisms was observed in the spleens and lungs determined over a 21-day period compared with age-matched normal controls. Mice immunized with the same culture filtrate proteins bound to nitrocellulose particles also showed some resistance to the virulent challenge, suggesting that individual antigens present in the culture filtrate were able to induce a protective T cell-mediated immune response in appropriately immunized mice.