男性不育患者精浆抗弓形虫IgG抗体与精子抗体关系的研究

目的:观察男性不育患者精浆中抗弓形虫 IgG 抗体。及其与抗精子抗体(AsAb)的关系。方法:采用 ELISA 方法测定169例男性不育患者及35例正常生育男性精浆抗弓形虫 IgG 抗体和 AsAb。结果:男性不育患者精浆抗弓形虫 IgG 抗体阳性率18.35%,显著高于正常生育组的2.86%(x~2=5.26,P<0.05)OR 为7.64;男性不育组精浆 AsAb 抗体阳性率为26.63%,也显著高于正常生育组(x~2=11.96,P<0.01)。精浆抗弓形虫抗体阳性的男性不育患者精浆AsAb 的阳性率为41.94%,与精浆抗弓形虫抗体阴性组的男性不育病人精浆 AsAb 的阳性率23.19%相比较,差异具有显著性(x~2=4.55,P<0.05).OR 为2.39。结论:男性不育患者精浆抗弓形虫抗体与精浆 AsAb 的产生可能有着一定的关系。     

Immunological approach to inhibit formation of anti-antibodies to allo- and xenogeneic anti-T cell immunoglobulin

Inhibitory anti-antibodies induced in patients by xenogeneic or even by humanized anti-T cell antibodies remain an unresolved problem. Mice also produce anti-antibodies following injection of xeno- or allogeneic anti-T cell antibodies. Here we report a principle based on sequentially applied anti-T cell antibodies generated in different species, which results in suppressed anti-antibody formation and prolonged immunosuppression. Thus, a single priming injection in mice of mouse (MmT1 or MmT5 differing by idiotype only) or of rat (RmT1) anti-mouse Thy-1 monoclonal antibodies (mAb) or of rat anti-mouse L3T4 + Ly-2 (RmCD4 + CD8) mAb suppressed anti-antibody formation against subsequent booster injections of one of the above antibodies, provided that they differed in species origin from the priming antibody. Correspondingly, a sixfold and longer prolongation of 50% survival of fully mismatched skin grafts was observed. Less or no anti-antibody suppression and little prolongation of graft surival was obtained if the ‘first’ and the ‘second’ (and following) antibody injections were of the same species, differing by iso- or idiotype only. Finally, the suppressive principle did not manifest itself at all if the initial antibody injection included both the first and second antibody. These findings are discussed with reference to earlier studies on hapten/carrier effects as well as on immunosuppression attributed to ‘non-depleting’ rat anti-CD4/CD8 T cell antibodies.     

Antibody-mediated activation of T cell clones as a method for screening hybridomas producing antibodies to the T cell receptor

In this study, supernatants (SN) of hybridomas established by fusing P3X63.Ag8.653 to spleen cells from C57L mice (Vß8 family of T cell receptor (TcR) gene negative) immunized with the H-Y specific Vß8 allotype positive helper T cell (HTL) clone OI6 were screened for the capacity to activate cloned T cells in the absence of interacting stimulator cells. In the first assay, SNs were mixed with Vß8⁺ H-Y specific CTL OH2 and ⁵¹Cr-labelled non-specific B lymphoma (L10.A). In this system, antibodies (Ab) which can bind to L10.A by Fc-Fc receptor interaction and recognize TcR can facilitate lysis of L10.A target cells by OH2 CTL. In the second assay, OI6 clone cells were cultured in microtiter well, previously coated with hybridoma supernatants (SN). In this assay, Ab recognizing OI6 TcR complex and bound to plastic plates can stimulate OI6 cells to proliferate in the absence of stimulator cells. Using these two screening methods, nine hybridomas were established. Analysis of these hybridoma SN using surface staining, inhibition of T cell function and immunoprecipitation of radiolabelled surface molecules followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS/PAGE) showed that five Ab were directed to the allotypic determinant (Vß8) of TcR and four Ab were specific to the clonotypic determinant of OI6 TcR. These results suggest that this Ab-mediated activation of T cell clone can be used for the screening of hybridomas secreting anti-TcR Ab and the immunogenicity of OI6 clonotypic determinants is similar to that of the Vß8 allotypic determinant even in strains which do not express the Vß8 TcR allotype.     

Patients' immune response to breast and lung carcinoma-associated thomsen-friedenreich (T) specificity

Summary We report here sensitive and specific measurement of immune responses of patients with certain kinds of carcinoma toward the physically and chemically well defined T antigen isolated from healthy human erythrocytes. Over 90% of adenocarcinoma tissues tested possess T-specific immunoreactive structures as determined withhuman antisera, in contrast to healthy tissues and benign lesions. Adenocarcinoma patients recognize the carcinoma-associated T antigen as foreign. Delayed-type skin hypersensitivity reaction to T antigen (DTHR-T) was positive in all 25 lung adenocarcinoma patients tested, in 88% of 101 patients with ductal, in 43% of 30 patients with lobular or tubular breast carcinoma and in 9/9 patients with adenocarcinoma of body cavities. Patients of all Stages reacted positively. All 7 patients with small cell lung carcinoma and 3/5 with malignant melanoma had a positive DTHR-T. None of 17 patients with malignant brain tumors, leukemia or Hodgkin's disease, sarcoma or thyroid carcinoma reacted. The DTHR-T was specific in that all 77 healthy persons and 48/49 with other diseases, including 23/24 with non-cancer lung disease were negative; one patient with organizing interstitial pneumonitis was positive. This points to a possible source of false positive reactions. 91% of 149 patients with histologically benign breast disease had a negative DTHR-T; the histology of some of the positive ones was reexamined, 2 proved to have carcinoma in situ. In vitro leukocyte migration inhibition and scoring of anti-T hemagglutinin titer using the T-anti-T system diagnosed many adenocarcinomata correctly, including 4 whose histology, while turning positive later, was negative at the time. However, these tests were generally less sensitive and specific than the DTHR-T.This system may also be of screening and monitoring value. Surgical removal of primary carcinoma led to a rebound of anti-T in breast carcinoma patients and its renewed decrease in some, prior to clinical recurrence of cancer. Also, the DTHR-T turned from positive to negative in some Stages I and II breast-and T_1–2 N_0–1 M₀ lung adenocarcinoma patients who had no demonstrable relapse during the ensuing observation period.Supported by U.S. Public Health Service Grants CA 19083 and CA 22540, American Cancer Society Research Development Program Grant RD-133, a gift from the Mobil fndn. and Cancer Res. Inst. N.Y. G.F.S. is Julia S. Michels Investigator in Surgical Oncology     

Effects of two doses of anti-T lymphocyte globulin-Fresenius given after full-match sibling stem cell transplantation in acute myeloblastic leukemia patients who underwent myeloablative fludarabine/busulfan conditioning

Objective/background

Anti-T lymphocyte globulin Fresenius (rATG-F; ATG-Fresenius) and antithymocyte globulin (thymoglobulin), which are included in transplant protocols, are used to reduce the risk of chronic graft-versus-host disease (cGVHD) or suppress allograft rejection. Available clinical studies have been conducted in heterogenous patient populations and with different administration protocols including stem cell sources. Additionally, the pharmacokinetics of ATG is variable, and the clinically effective dose of rATG-F, in particular, is not exactly known. The aim of the study was to investigate the clinical outcomes of acute myeloid leukemia (AML) patients who underwent hemopoietic peripheral stem cell transplantation from full-matched sibling donors and given two different doses of r-ATG-F.

Immunosuppression by Fc region-mismatched anti-T cell antibody treatment

Formation of anti-antibodies (anti-Ab) is known to counteract immunotherapy with anti-T cell antibodies. Our previously described immunological approach prevented anti-Ab with the consequence of prolonged survival of fully mismatched skin grafts in C57BL/6 mice. These mice were treated with a single priming injection of a monoclonal anti-T cell Ab followed by repeated injections of anti-T cell mAb differing in species origin from the priming mAb. We now show prolonged tolerance to discordant xenogeneic, to bispecific, and even to polyclonal Ab, and demonstrate that the underlying immunosuppressive principle is due to a difference in heavy chain constant region between first and second antibodies, independent of whether or not they share the same idiotype. To examine this phenomenon, a panel of mAb was generated which share the same mouse anti-Thy-1.2 idiotype, but carry a human IgG1(T23), IgG3(T212C8), or mouse IgG2a(MmT1) constant heavy chain region. We found that sequential injection of MmT1 and T23 according to the above treatment schedule induced huIgG1 isotype-specific tolerance to T23, which was similar to that seen when using a primary mAb (MmT5) that was, instead, fully mismatched with T23 in both idiotype and constant region. Thus, differences of idiotype between primary and booster Ab were inconsequential for their ability to inhibit anti-Ab formation. This novel form of induced specific tolerance to anti-T cell Ig survived graft rejection and was still evident 230 days after termination of the T cell depletion protocol. Taken together, these results demonstrate that rechallenge with Fc region-mismatched Ab opens an immunological window that allows for induction of tolerance to immunogenic anti-T cell Ab and prolonged immunosuppression.     

Anti-T antibodies and peanut-agglutinin-binding glycoproteins in sera of patients with gastric cancer

Agglutinating antibodies to neuraminidase-treated red blood cells (anti-T agglutinins) are known to be reduced in patients with gastric cancer. The antigenic determinant of anti-T agglutinin is known to have a disaccharide structure [Gal(β1-3)GalNAc], the same specificity as peanut agglutinin (PNA). We examined sera of 27 patients with gastric cancer and 30 controls for anti-T agglutinins, anti-T antibodies and PNA-binding glycoproteins. Anti-T agglutinins were titrated by a microtiter hemagglutination method. Levels of anti-T antibodies were determined by enzyme immunoassay using synthetic glycoconjugate [Gal(β1-3)GalNAc O-α-linked to human serum albumin] as an antigen. Levels of PNA-binding glycoproteins in sera were measured by sandwich enzyme-linked lectin assay using wheat germ agglutinin and peroxidase-conjugated PNA. Titers of anti-T agglutinins were significantly lower in patients with gastric cancer than in controls (P = 0.041). Levels of anti-T antibodies were not significantly different in patients with gastric cancer and controls; however, decreased levels of anti-T antibodies were more frequent in patients with gastric cancer than in controls (P = 0.001). Levels of PNA-binding glycoproteins were significantly higher in sera of patients with gastric cancer than in controls (P = 0.001). The levels of anti-T antibodies inversely correlated with the levels of PNA-binding glycoproteins in sera of patients with gastric cancer (r = −0.44, P = 0.021). These results suggest that the decrease in anti-T antibodies in sera of patients with gastric cancer might be due to immune complex formation between circulating PNA-binding glycoproteins and anti-T antibodies. Received: 3 March 1999 / Accepted: 19 May 1999     

男性不育精浆弓形虫IgG与NO关系研究

目的　观察男性不育病人精浆弓形虫IgG与NO水平及其关系。 方法　采用ELISA方法测定 16 9例男性不育患者及 35例正常生育男性精浆弓形虫IgG抗体 (ATAb)和抗精子抗体 (AsAb) ,同时采用硝酸还原酶法测定精浆中NO水平。结果　男性不育病人精浆ATAb阳性率 18 35 % ,显著高于正常生育组的 2 86 % (P <0 0 5 ) ;男性不育组精浆AsAb阳性率为2 6 6 3% ,也显著高于正常生育组 (P <0 0 1)。男性不育组精浆中NO水平 14 6 6 8± 38 87μmol/L ,显著高于正常生育组 84 92± 2 6 72 μmol/L(P <0 0 1)。精浆弓形虫IgG抗体阳性的男性不育病人精浆NO水平 15 3 2 1± 4 1 6 0 μmol/L ,显著高于精浆ATAb阴性组的男性不育病人精浆的NO水平 135 81± 31 4 5 μmol/L(P <0 0 1)。精浆AsAb阳性的男性不育病人NO水平15 8 76± 4 3 0 0 μmol/L ,显著高于精浆AsAb阴性组的男性不育病人精浆的 133 82± 32 10 μmol/L(P <0 0 1)。精浆ATAb阳性的男性不育病人精浆AsAb的阳性率为 38 71% ,也显著高于精浆ATAb阴性组的男性不育病人精浆AsAb的阳性率2 3 91% (P <0 0 1)。结论　男性不育病人精浆ATAb与精浆NO水平具有相关性 ,同时也与抗精子抗体的产生可能有着一定的关系。     

Thomsen-Friedenreich antigen in bladder tumors as detected by specific antibody: A possible marker of recurrence

Summary TUR specimens of non-invasive transitional-cell carcinomas were examined for their expression of Thomsen-Friedenreich (T) antigen by indirect immunofluorescence staining using rabbit IgG antibody which was raised with desialated glycophorin. Nine (45%) out of 20 tumors of low grade (Grade I and II), and 5 (56%) of 9 tumors of high grade (III) were diffusely stained with anti-T (T-positive), whereas T-antigen in normal tissue was cryptic and stained only after neuraminidase treatment (cryptic T-positive). Of the T-negative tumors, 9 (45%) of the low grade but only one of the high grade tumors, were stained positively for the cryptic T-antigen. The rest of the tumors were devoid of the cryptic T-antigen. Eighty percent of both Tumors expressing T-antigen and those lacking the cryptic T-antigen recurred within two years. Recurrence was not influenced by initial histological grade.     

Murine anti-mouse T cell monoclonal antibodies elicit anti-antibodies in mice: intra-species immunization model for estimating potential patient sensitization against humanized anti-T cell antibodies

Humanization of immunosuppressive anti-T cell monoclonal antibodies (mAb) raises the question as to how completely it helps to avoid formation of neutralizing anti-antibodies (anti-Ab) in patients. To get more information on intra-species sensitization against anti-T cell mAb, we produced two immunosuppressive mouse IgG2a anti-mouse Thy-1.2 mAb (MmT1 and MmT5) in AKR/J mice and measured the potential of MmT1 to elicit inhibitory anti-Ab in AKR/J (H-2^k), C57BL/6 (H-2^b), congenic B10.BR (H-2^k) and DBA/2 (H-2^d) mice. After one injection once weekly for 4 weeks of 5 μg MmT1 (200 μg/kg) in C57BL/6 mice, without the use of any adjuvants, high concentrations of anti-Ab directed against MmT1 (300 μg/ml) and MmT5 (100 μg/ml) were measured by enzyme-linked immunosorbent assay. Similar concentrations of anti-Ab were found in immunized DBA/2 and less in B10.BR mice. No syngeneic anti-Ab could be produced in AKR/J. From the C57BL/6 mice, we raised anti-MmT1⁺, MmT5⁺ idiotype (IDIO1) and anti-MmT1⁺, MmT5⁺ allotype (ALLO1) mAb. An in vivo test system was adapted to measure the inhibitory effects of circulating poly- or monoclonal anti-Ab. It revealed a reduction of in vivo depletion capacity not only of the sensitizing mAb (MmT1), but also of another anti-Thy-1.2 mAb (MmT5), with identical allotype but different idiotype. From this we conclude that intra-species immunization following injection of anti-T cell mAb can produce high titer inhibitory anti-idiotype and anti-allotype antibodies. Implications for hyperchimeric or fully human anti-T cell mAb are discussed.