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1.
PLoS One ; 8(10): e76839, 2013.
Article in English | MEDLINE | ID: mdl-24194847

ABSTRACT

Immune suppression with rabbit anti-thymocyte globulin (rATG) is a well-established therapeutic concept for preventing host rejection of transplanted organs and graft versus host disease. Increasing the efficiency of rATG production by reducing the number of animals would be highly beneficial to lower cost and to improve quality standards. We have developed transgenic (Tg) mice and rabbits that overexpress the neonatal Fc receptor (FcRn) and have shown an augmented humoral immune response in these animals. To test whether our FcRn Tg rabbits produced rATG more efficiently, we immunized them and their New Zealand White controls with live Jurkat cells. By day 21 after immunization, Tg animals produced significantly, 1.5 times higher amount of total IgG compared to their wt littermates. Also, the binding efficiency of Tg sera to Jurkat cells and their complement-mediated cytotoxicity was significantly higher. The purified Tg IgG preparation contained 2.6 the amount of Jurkat specific IgG as the wt preparation analyzed by complement-mediated lysis, suggesting greater antigen-specific B cell activation in the Tg rabbits. To test this hypothesis, immunization with ovalbumin and human α1-antitrypsin was performed, resulting in significantly greater numbers of antigen-specific B-cells in the FcRn Tg rabbits as compared with wt controls. The shift towards significantly larger populations of antigen-specific B cells relative to the non-specific B cell pool is further corroborated by our previous findings in FcRn Tg mice. Consequently, our FcRn Tg rabbits have the potential to offer substantial qualitative and quantitative improvements for the production of rATG and other polyclonal or monoclonal antibodies.


Subject(s)
Animals, Genetically Modified/immunology , Antilymphocyte Serum/biosynthesis , B-Lymphocytes/immunology , Graft Rejection/prevention & control , Histocompatibility Antigens Class I/metabolism , Rabbits/immunology , Receptors, Fc/metabolism , Animals , Antilymphocyte Serum/therapeutic use , B-Lymphocytes/drug effects , Chromatography, Agarose , Enzyme-Linked Immunosorbent Assay , Enzyme-Linked Immunospot Assay , Flow Cytometry , Histocompatibility Antigens Class I/immunology , Humans , Immunoglobulin G/biosynthesis , Jurkat Cells , Lymphocyte Activation/drug effects , Mice , Ovalbumin/pharmacology , Receptors, Fc/immunology , alpha 1-Antitrypsin/pharmacology
2.
Transplant Proc ; 45(4): 1528-30, 2013 May.
Article in English | MEDLINE | ID: mdl-23726612

ABSTRACT

Recipients of primary transplants from donation after cardiac death (DCD) donors (n = 40) performed from January 2005 to December 2009 were retrospectively reviewed and compared with recipients of primary transplants from donation after brain death (DBD) donors (n = 142). Patients received rabbit antithymocyte globulin induction and rapid steroid taper (RST; steroids stopped 5 days after surgery). Maintenance immunosuppression included tacrolimus and mycophenolate mofetil. Protocol kidney biopsies, creatinine (Cr), and measured glomerular filtration rate (mGFR; determined by cold iothalamate or 24-h creatinine clearance) were obtained at 1, 4, 12, and 24 months. Kidney biopsies for cause were conducted for unexplained elevated Cr, decline in mGFR, or new proteinuria. Biopsies were graded for rejection according to the Banff criteria. Graft survival at 3 years was 90.0% for DCD recipients and 86.6% for DBD recipients (P = NS). Rejection of any grade diagnosed on any biopsy through the first 2 years occurred in 18 DCD (45%) and 50 DBD (35%) recipients. Rejection of a grade more than Banff borderline occurred in 12.5% DCD and 12.7% DBD recipients. At 2 years, the mean ± SEM Cr and mGFR for DCD recipients with rejection were 1.8 ± 0.29 mg/dL and 59.2 ± 8.5 mL/min versus 1.3 ± 0.11 mg/dL and 67.0 ± 7.8 ml/min for those without rejection. For DBD recipients with rejection, Cr and mGFR at 2 years were 1.7 ± 0.12 mg/dL and 54.0 ± 4.4 mL/min versus 1.4 ± 0.11 mg/dL and 66.6 ± 3.3 ml/min for those without rejection (P = NS). Comparing DCD to DBD, there was no statistical difference in mean Cr or mGFR outcomes. Regardless of group, grafts with delayed graft function had lower 3-year survival. DCD primary kidney transplant recipients treated with rabbit antithymocyte induction and RST have short-term graft survival and function equivalent to DBD recipients. RST appears to be acceptable immunosuppression for DCD recipients.


Subject(s)
Antilymphocyte Serum/biosynthesis , Death , Delayed Graft Function , Graft Rejection , Kidney Transplantation , Steroids/administration & dosage , Tissue Donors , Aged , Creatinine/blood , Female , Glomerular Filtration Rate , Humans , Male , Middle Aged
3.
Fitoterapia ; 77(2): 100-8, 2006 Feb.
Article in English | MEDLINE | ID: mdl-16376495

ABSTRACT

A competitive enzyme-linked immunosorbent assay (ELISA) for saikosaponins was established using monoclonal antibody (MAb) 3G10. Hybridoma 3G10 prepared by fusing splenocytes immunized with saikosaponin a-BSA (SSa-BSA) conjugate and a hypoxanthine-aminopterin-thymidine (HAT)-sensitive mouse myeloma cell line, P3-X63-Ag8-U1, secreted monoclonal antibodies with wide cross-reactivity to saikosaponins including saikosaponin b(2) (SSb(2)), c (SSc) and d (SSd), which are stereo and/or regio isomers of SSa. The method, at an effective measuring range of 0.6 mug /ml to 2.3 mug/ml of SSa, successfully detected total saikosaponins in Bupleuri radix and Kampo medicines prescribed with Bupleuri radix. Good correlation between ELISA and HPLC analyses of total saikosaponin in a crude extract of Bupleuri radix was obtained after hydrolysis of acyl saikosaponins by treatment with a mild alkaline solution.


Subject(s)
Drugs, Chinese Herbal/analysis , Enzyme-Linked Immunosorbent Assay/methods , Oleanolic Acid/analogs & derivatives , Plant Extracts/analysis , Saponins/analysis , Animals , Antibodies, Monoclonal/immunology , Antilymphocyte Serum/analysis , Antilymphocyte Serum/biosynthesis , Bupleurum/chemistry , Calibration , Cell Line, Tumor , Chromatography, High Pressure Liquid/methods , Cross Reactions , Female , Haptens/analysis , Hybridomas , Linear Models , Mice , Mice, Inbred BALB C , Oleanolic Acid/analysis , Oleanolic Acid/isolation & purification , Plant Extracts/chemistry , Plants, Medicinal/chemistry , Saponins/isolation & purification , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Statistics as Topic/methods
4.
Springer Semin Immunopathol ; 26(4): 433-52, 2005 Mar.
Article in English | MEDLINE | ID: mdl-15633016

ABSTRACT

B cells play a variety of immunoregulatory roles through their antigen-presentation ability and through cytokine and chemokine production. Innate immune activation of B cells may play a beneficial role through the generation of natural cross-reactive antibodies, by maintaining B cell memory and by exercising immunomodulatory functions that may provide protection against autoimmunity. In this article, we review human B cell populations and their functional properties, with a particular focus on a population of inherently autoreactive B cells, which seem to play an important physiological role in innate immunity, but which, if selected into adaptive immune responses, appear to become pathogenic agents in systemic lupus erythematosus.


Subject(s)
Autoimmunity , B-Lymphocyte Subsets/immunology , Immunity, Innate , Antilymphocyte Serum/biosynthesis , B-Lymphocyte Subsets/cytology , Cell Differentiation , Humans , Lupus Erythematosus, Systemic/immunology , Models, Immunological
5.
Springer Semin Immunopathol ; 26(4): 363-75, 2005 Mar.
Article in English | MEDLINE | ID: mdl-15611857

ABSTRACT

Natural antibodies produced by CD5+ B-1 B cells include those with specificity for senescent erythrocytes (anti-BrMRBC, anti-PtC) and for thymocytes (anti-thymocyte autoantibody, ATA). Here we describe work from our laboratories studying two prototypic examples, V(H)11Vkappa9-encoded anti-BrMRBC and V(H)3609Vkappa21c-encoded ATA. Using V(H)11-mu transgenic mice, we discovered that certain natural autoantibodies utilize V(H) genes that are selected against in bone marrow B cell development, but not fetal liver, effectively restricting their generation to fetal/neonatal life. Studies with ATA-mu transgenic mice demonstrated a critical requirement for self antigen in the accumulation of B cells with this specificity and for the production of high levels of serum ATA. Finally, analysis of B cell development in ATA-mu kappa transgenic mice revealed two distinct responses by B cells to expression of this B cell receptor (BCR): most developing B cells in spleen of adult mice were blocked at an immature stage and only escaped apoptosis by editing their BCR to eliminate the ATA specificity; nevertheless, high levels of serum ATA were observed, indicating that some B cells differentiated to antibody-forming cells without altering their specificity. Thus, our studies reveal mechanisms for restricting the generation of B cells producing natural autoantibodies, demonstrate a key positive selection step in their development, and show that most developing B cells in adult mice bearing such specificities fail to reach a mature stage.


Subject(s)
B-Lymphocyte Subsets/immunology , Immunity, Innate , Animals , Antibody Specificity , Antilymphocyte Serum/biosynthesis , Autoantibodies/biosynthesis , B-Lymphocyte Subsets/cytology , Cell Differentiation , Erythrocyte Aging , Erythrocytes/immunology , Mice , Mice, Transgenic , Models, Immunological , Receptors, Antigen, B-Cell/metabolism , T-Lymphocytes/immunology
6.
J Am Soc Nephrol ; 13(3): 769-772, 2002 Mar.
Article in English | MEDLINE | ID: mdl-11856783

ABSTRACT

A total of 73,707 primary renal transplants reported to the USRDS between 1988 and 1997 were examined to investigate the cause-specific risk for patient death associated with anti-lymphocyte antibody induction therapy (ABI). Cox proportional hazard models were used to estimate the relative risk of the use of ABI and patient death. All Cox models were corrected for potential confounding variables, such as age, gender, race, HLA mismatch, panel reactive antibody, delayed graft function, cold ischemia time, time since start of dialysis, etiology of end-stage renal disease, cytomegalovirus risk group, donor source (living or cadaveric), era effect, and immunosuppressive therapy. Primary study end points were patient death with functioning graft (DWFG) and overall patient death, including death after graft loss. Early patient death (deaths within the first 6 mo after renal transplantation) and late death (deaths after 6 mo post-renal transplantation) were investigated separately. Additionally, specific causes of death were investigated. ABI was associated with a significant risk for late death after renal transplantation (relative risk [RR] = 1.1; P < 0.001) but not for DWFG (RR = 0.94; P = 0.10). ABI conferred the highest RR for late malignancy-related death (RR = 1.35; P < 0.001). ABI was significantly associated with early deaths due to infection and cardiovascular causes (RR = 1.32 [P < 0.001] and RR = 1.27 [P < 0.001], respectively). Kaplan Meier plots confirmed that the risk of ABI for patient death secondary to infectious complications was increased predominately early after transplantation as opposed to late for malignancy-related death. ABI was associated with a significant relative risk for patient death secondary to cardiovascular causes and infectious complications early in the posttransplant period. In addition, ABI was associated with a significant risk for long-term malignancy-related death. The risk of ABI should be taken in context with potential benefits of this therapy.


Subject(s)
Antilymphocyte Serum/biosynthesis , Kidney Transplantation/immunology , Kidney Transplantation/mortality , Muromonab-CD3/adverse effects , Adult , Cause of Death , Female , Humans , Infections/etiology , Infections/mortality , Kidney Transplantation/adverse effects , Male , Middle Aged , Multivariate Analysis , Proportional Hazards Models , Risk Factors , Survival Analysis
7.
J Immunol ; 166(12): 7072-81, 2001 Jun 15.
Article in English | MEDLINE | ID: mdl-11390451

ABSTRACT

We studied the effect of host IFN-gamma on the pathology of acute rejection of vascularized mouse heart and kidney allografts. Organs from CBA donors (H-2k) were transplanted into BALB/c (H-2d) hosts with wild-type (WT) or disrupted (GKO, BALB/c mice with disrupted IFN-gamma genes) IFN-gamma genes. In WT hosts, rejecting hearts and kidneys showed mononuclear cell infiltration, intense induction of donor MHC products, but little parenchymal necrosis at day 7. Rejecting allografts in GKO recipients showed infiltrate but little or no induction of donor MHC and developed extensive necrosis despite patent large vessels. The necrosis was immunologically mediated, since it developed during rejection, was absent in isografts, and was prevented by immunosuppressing the recipient with cyclosporine or mycophenolate mofetil. Rejecting kidneys in GKO hosts showed increased mRNA for heme oxygenase 1, and decreased mRNA for NO synthase 2 and monokine inducible by IFN-gamma (MIG). The mRNA levels for CTL genes (perforin, granzyme B, and Fas ligand) were similar in rejecting kidneys in WT and GKO hosts, and the host Ab responses were similar. The administration of recombinant IFN-gamma to GKO hosts reduced but did not fully prevent the effects of IFN-gamma deficiency: MHC was induced, but the prevention of necrosis and induction of MIG were incomplete compared with WT hosts. Thus, IFN-gamma has unique effects in vascularized allografts, including induction of MHC and MIG, and protection against parenchymal necrosis, probably at the level of the microcirculation. This is probably a local action of IFN-gamma produced in large quantities in the allograft.


Subject(s)
Graft Rejection/immunology , Graft Rejection/pathology , Heart Transplantation/immunology , Heart Transplantation/pathology , Interferon-gamma/physiology , Kidney Transplantation/immunology , Kidney Transplantation/pathology , Acute Disease , Animals , Antibody-Dependent Cell Cytotoxicity/genetics , Antilymphocyte Serum/biosynthesis , Gene Expression Regulation/genetics , Gene Expression Regulation/immunology , Graft Rejection/genetics , Graft Rejection/prevention & control , Histocompatibility Antigens Class I/biosynthesis , Histocompatibility Antigens Class II/biosynthesis , Immune Sera/administration & dosage , Injections, Intraperitoneal , Interferon-gamma/administration & dosage , Interferon-gamma/genetics , Interferon-gamma/immunology , Kidney/blood supply , Kidney/immunology , Kidney/pathology , Male , Mice , Mice, Inbred BALB C , Mice, Inbred CBA , Mice, Knockout , Myocardium/immunology , Myocardium/pathology , Necrosis , Recombinant Proteins/administration & dosage
8.
J Immunol ; 165(2): 637-44, 2000 Jul 15.
Article in English | MEDLINE | ID: mdl-10878335

ABSTRACT

The association of preformed anti-donor Abs with the hyperacute rejection of bone marrow and solid organ allografts and the persistence of the anti-donor immune response secondary to immunologic memory make allosensitization an absolute contraindication to transplantation. Mixed allogeneic (A + B-->A) bone marrow chimerism has been demonstrated to confer donor-specific tolerance in nonsensitized recipients, but has not been evaluated in the setting of allosensitization. The current study documents that despite significant anti-donor sensitization, mixed allogeneic engraftment is possible and provides a marked advantage over fully allogeneic (B-->A) models. Moreover, the acceptance of donor skin grafts and loss of circulating anti-donor Abs suggest that allosensitization can be abrogated with the induction of stable mixed allogeneic chimerism.


Subject(s)
Graft Rejection/genetics , Graft Rejection/immunology , Immunization , Isoantigens/immunology , Radiation Chimera/immunology , Animals , Antibody Specificity/genetics , Antilymphocyte Serum/biosynthesis , Bone Marrow Transplantation/immunology , Cytotoxicity, Immunologic/genetics , Graft Survival/genetics , Graft Survival/immunology , Immune Tolerance/genetics , Immunization/methods , Immunologic Memory/genetics , Isoantigens/genetics , Isoantigens/radiation effects , Kinetics , Male , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Skin Transplantation/immunology
9.
Transpl Immunol ; 8(3): 203-9, 2000 Nov.
Article in English | MEDLINE | ID: mdl-11147701

ABSTRACT

To compare the effectiveness of transgenic and systemic monoclonal antibody therapy for pancreas transplantation, vascularised segmental pancreas allografts from wild-type or transgenic pancreatic tissue that secreted monoclonal anti-CD4 were placed in CBA recipients in which diabetes had been induced chemically by streptozotocin (STZ, non-autoimmune diabetes). In untreated CBA recipients, wild-type BALB/c or C57BL/6 bml pancreas transplants were rejected in a mean survival time (MST) of 27 and 30 days, respectively. BALB/c and C57BL/6 graft survival improved when recipients were given a short course of T cell depleting monoclonal anti-CD4 antibody, (GK 1.5, 2 mg total on days -1, 0, 1, 2 with grafting on day 0) with MST +/- S.D. of 71 +/- 29 and 44 +/- 36 days, respectively. Thus, transient depletion of CD4 was effective in delaying pancreas allograft rejection in these strain combinations. The use of C57BL/6 bml mice transgenic for a rat anti-CD4 antibody (GK5 mice) as pancreas donors provided allografts that secreted sufficient anti-CD4 antibody to cause CD4 T cell depletion in the recipients (CD4 cells decreased from 30 to < 5% of small lymphocytes). This degree of depletion was not sustained and the CD4 recovery inversely correlated with graft survival. Mice with > 20% CD4 cells in the splenic lymphocyte population 4 weeks post-transplant rejected their grafts (3 of 10 mice). However, in 7 of 10 mice CD4 cells remained low (< 15%) and allografts survived for > 80 days. The GK5 allografts survived significantly longer than those from non-transgenic bml controls (MST 83 +/- 32 days, compared with 30 days, P < 0.0005). This survival time was similar to that of BALB/c allografts in CBA recipients treated with a high dose of anti-CD4 antibody. Thus, transgenic secretion of anti-CD4 antibody by the pancreas allograft was very effective in prolonging its survival.


Subject(s)
Antibodies, Monoclonal/biosynthesis , CD4 Antigens/immunology , Graft Enhancement, Immunologic/methods , Graft Survival/immunology , Pancreas Transplantation/immunology , Animals , Antibodies, Monoclonal/genetics , Antilymphocyte Serum/biosynthesis , Antilymphocyte Serum/genetics , Diabetes Mellitus, Experimental/surgery , Graft Survival/genetics , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Inbred CBA , Mice, Transgenic , Pancreas Transplantation/pathology , Rats , Recombinant Proteins/biosynthesis , Recombinant Proteins/genetics , T-Lymphocytes/immunology , Time Factors , Transplantation Immunology , Transplantation, Homologous
10.
J Periodontal Res ; 34(2): 105-12, 1999 Feb.
Article in English | MEDLINE | ID: mdl-10207839

ABSTRACT

We examined the levels of anti-thymocyte/T lymphocyte autoantibody (ATA) in the serum of mice injected intraperitoneally with lipopolysaccharides (LPS) from periodontopathic bacteria; Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Fusobacterium nucleatum, Capnocytophaga ochracea, and non-oral Escherichia coli. All of the LPS induced IgM-ATA. Among these, LPS from C. ochracea induced the highest level of IgM-ATA, whereas that of P. gingivalis induced the lowest. The peritoneal T lymphocytes of mice injected with LPS were bound by IgM-ATA. Peritoneal B-1 (CD5+B) cells stimulated by each LPS produced much more IgM-ATA than splenic B-2 (CD5-B) cells, suggesting that B-1 cells might be responsible for the production of these antibodies. Serum of mice injected with C. ochracea and F. nucleatum LPS showed cytotoxicity against thymocytes in the presence of rabbit complements. Binding and cytotoxicity were confirmed by IgM purified from serum of the mice injected with C. ochracea LPS. Furthermore, serum of mice treated with C. ochracea, F. nucleatum or A. actinomycetemcomitans LPS inhibited the proliferation of thymocytes. However, purified IgM from the serum of mice treated with C. ochracea LPS failed to produce the same inhibition. Our results suggest that LPS from certain species of periodontopathic bacteria can induce IgM-ATA in the serum and these antibodies may modulate the local immune network in periodontal tissues.


Subject(s)
Autoantibodies/biosynthesis , Immunoglobulin M/biosynthesis , Lipopolysaccharides/pharmacology , Periodontitis/microbiology , T-Lymphocytes/drug effects , Aggregatibacter actinomycetemcomitans/immunology , Analysis of Variance , Animals , Antilymphocyte Serum/biosynthesis , Autoantibodies/blood , B-Lymphocytes/drug effects , Capnocytophaga/immunology , Cytotoxicity, Immunologic , Escherichia coli/immunology , Fusobacterium nucleatum/immunology , Immunoglobulin M/blood , Male , Mice , Mice, Inbred BALB C , Mice, Inbred NZB , Periodontitis/immunology , Porphyromonas gingivalis/immunology , Rabbits , Statistics, Nonparametric
11.
J Autoimmun ; 10(2): 137-46, 1997 Apr.
Article in English | MEDLINE | ID: mdl-9185875

ABSTRACT

T cell vaccination, the application of syngeneic attenuated T cells, has been shown to prevent effectively and treat experimental autoimmune diseases, but its mechanisms of action are poorly understood. Here we present data on the induction of a humoral anti-T cell response by T cell vaccination, capable of strongly inhibiting T cell proliferation and of ameliorating experimental autoimmune disease. T cell vaccination in the Lewis rat induced autoantibodies reactive with several syngeneic T cell proteins. These autoantibodies were not detectable in normal Lewis sera as assessed by immunoblotting and flow cytometry with intact syngeneic T cells. The autoantibody reactivity was not restricted to one idiotype, was detected as early as 1 week after vaccination and was dominated by IgG, suggesting the boosting of a naturally preformed humoral network by T cell vaccination. Recovery from passively or actively induced experimental autoimmune encephalomyelitis (EAE) in the Lewis rat, too, could be shown to be associated with the development of anti-T cell autoantibodies. In vitro, both the post-EAE and the post-vaccination sera had a strong suppressive effect on the proliferation of syngeneic T cell clones. This inhibition was shown to be mediated by antibodies and to be partly complement-dependent. In vivo, both kinds of sera were able to ameliorate EAE. This protective effect of the post-vaccination sera was not idiotype-specific, since sera obtained after T cell vaccination with an unrelated T cell clone were similarly effective in suppressing EAE. These results suggest that anti-lymphocytic antibodies might play an immunoregulatory role that can be positively manipulated by T cell vaccination.


Subject(s)
Antilymphocyte Serum/biosynthesis , Antilymphocyte Serum/chemistry , Autoantibodies/biosynthesis , Autoantibodies/chemistry , T-Lymphocytes/transplantation , Vaccination , Animals , Antilymphocyte Serum/physiology , Autoantibodies/physiology , Clone Cells/transplantation , Encephalomyelitis, Autoimmune, Experimental/blood , Encephalomyelitis, Autoimmune, Experimental/immunology , Encephalomyelitis, Autoimmune, Experimental/therapy , Immune Sera/pharmacology , Immunoglobulin Idiotypes/biosynthesis , Immunoglobulin Idiotypes/chemistry , Lymphocyte Activation , Membrane Proteins/immunology , Rats , Rats, Inbred Lew
12.
J Immunol ; 158(6): 2695-703, 1997 Mar 15.
Article in English | MEDLINE | ID: mdl-9058803

ABSTRACT

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD), a highly toxic environmental contaminant, suppresses both CTL and cytotoxic alloantibody production in C57BL/6 mice challenged with allogeneic P815 tumor cells. Recent evidence suggests that TCDD interferes with the initial activation of CD4+ Th cells, possibly through an indirect mechanism. In this study, we examined the effect of TCDD on the expression of the important costimulatory molecules, B7-1 and B7-2, in P815 allograft immunity. Expression of B7-2, but not B7-1, was up-regulated on splenic B220+ and Mac-1+ cells in P815-challenged mice. Exposure to TCDD significantly decreased the expression of B7-2 on B220+ and Mac-1+ cells in P815-challenged mice. Providing exogenous B7-mediated costimulation, in the form of B7-transfected P815 tumor cells, induced CTL activity in TCDD-treated mice by a mechanism that was independent of CD4+ T cells. In contrast, B7-transfected P815 cells did not restore the cytotoxic alloantibody response in TCDD-treated mice. These results are consistent with a model in which MHC class II-, B7-transfected P815 tumor cells can directly activate CD8+ CTL precursors but cannot directly stimulate CD4+ T helper cells required for B cell activation. These results also demonstrate that CTL precursors in TCDD-treated mice are functional and able to differentiate into effector CTL provided they receive adequate costimulation via B7 and suggest that defective costimulation, through reduced B7-2 expression, may play a role in TCDD-induced immunotoxicity. In support of this hypothesis, we show that blocking B7-2/CD28 interactions, and to a lesser degree B7-1/CD28 interactions, suppressed the alloimmune responses to P815 tumor cells, which further indicates that B7-2 represents the dominant B7 molecule involved in the generation of an immune response to allogeneic P815 tumor cells.


Subject(s)
B7-1 Antigen/biosynthesis , B7-1 Antigen/genetics , Cytotoxicity, Immunologic/drug effects , Isoantibodies/biosynthesis , Mast-Cell Sarcoma/immunology , Polychlorinated Dibenzodioxins/toxicity , T-Lymphocytes, Cytotoxic/drug effects , T-Lymphocytes, Cytotoxic/immunology , Animals , Antigens, CD/biosynthesis , Antilymphocyte Serum/biosynthesis , Antilymphocyte Serum/drug effects , B7-1 Antigen/drug effects , B7-2 Antigen , CD4-Positive T-Lymphocytes/immunology , Cytotoxicity, Immunologic/genetics , Leukocyte Common Antigens/drug effects , Macrophage-1 Antigen/drug effects , Male , Mast-Cell Sarcoma/genetics , Membrane Glycoproteins/biosynthesis , Mice , Mice, Inbred C3H , Mice, Inbred C57BL , Mice, Inbred DBA , Neoplasm Transplantation , Spleen/cytology , Spleen/immunology , T-Lymphocytes, Cytotoxic/metabolism , Transfection , Tumor Cells, Cultured
13.
J Immunol ; 157(6): 2310-9, 1996 Sep 15.
Article in English | MEDLINE | ID: mdl-8805628

ABSTRACT

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD), a widespread environmental contaminant and prototypic ligand for the aryl hydrocarbon receptor, is a potent immunotoxicant. To understand the underlying mechanisms of TCDD immunotoxicity, we have characterized the time course of changes in CTL, alloantibody, and cytokine responses to the P815 tumor allograft in C57B1/6 mice treated with 0 or 15 microg TCDD/kg. Suppression of CTL activity by TCDD directly correlated with reduced numbers of splenic CTL effector cells identified by their CD8+CD44 high CD45RB low phenotype, while suppression of the alloantibody response correlated with a lack of expansion of the B220+ splenocyte population. Cytokine production was differentially modulated following TCDD treatment. Although type 1 cytokine production (IFN-gamma, IL-2, and TNF) was initially induced in TCDD-treated mice, production failed to increase normally after day 5. In contrast, the production of IL-1 beta, IL-4, and IL-6 was mostly unaffected by TCDD exposure. This differential effect of TCDD on cytokine production was reflected in the degree of suppression of specific alloantibody isotypes. TCDD abrogated the production of IgG2a (promoted by IFN-gamma), but had much less effect on the level of IgG1 (promoted by IL-4). IgM Ab titers were also highly suppressed. CD8+ cells were the exclusive producers of IFN-gamma and IL-2 when spleen cells from P815-injected mice were cultured in vitro on days 4 to 7 after P815 injection. However, CD4+ cells were shown to play a crucial role in the generation of both CTL and alloantibody responses, since their depletion in vivo abolished both responses. Based on similar temporal effects produced by TCDD and anti-CD4 Ab on alloimmune responses, we postulate that TCDD interferes with the initial activation of CD4+ T cells, which leads to downstream inhibition of the activation and/or differentiation of CD8+ T cells and B cells. In addition, since delayed treatment with either anti-CD4 Ab or TCDD suppressed the alloantibody but not the CTL response, TCDD may also affect later CD4+ T helper-B cell interactions.


Subject(s)
Antibody-Dependent Cell Cytotoxicity/drug effects , Cytokines/antagonists & inhibitors , Cytokines/biosynthesis , Immunosuppressive Agents/toxicity , Isoantibodies/biosynthesis , Isoantibodies/drug effects , Polychlorinated Dibenzodioxins/toxicity , T-Lymphocytes, Cytotoxic/drug effects , Animals , Antilymphocyte Serum/biosynthesis , CD4-Positive T-Lymphocytes/drug effects , CD8-Positive T-Lymphocytes/classification , CD8-Positive T-Lymphocytes/drug effects , Cytotoxicity Tests, Immunologic , Female , Immunoglobulin Isotypes/drug effects , Immunoglobulin Isotypes/immunology , Immunophenotyping , Interferon-gamma/biosynthesis , Interferon-gamma/drug effects , Interferon-gamma/genetics , Interleukin-2/biosynthesis , Interleukin-2/genetics , Lymphocyte Activation/drug effects , Lymphocyte Count/drug effects , Lymphocyte Depletion , Male , Mast-Cell Sarcoma , Mice , Mice, Inbred C57BL , Mice, Inbred DBA , Neoplasm Transplantation , RNA, Messenger/drug effects , Spleen/cytology , T-Lymphocytes, Cytotoxic/immunology , Tumor Cells, Cultured , Tumor Necrosis Factor-alpha/drug effects , Tumor Necrosis Factor-alpha/genetics
14.
Transpl Immunol ; 3(1): 39-44, 1995 Mar.
Article in English | MEDLINE | ID: mdl-7551977

ABSTRACT

The intrathymic inoculation (ITI) of donor splenocytes into potential organ transplant recipients has been demonstrated to result in donor-specific unresponsiveness and greatly prolonged survival of subsequent organ allografts in rodents without the need for long-term pharmacological immunosuppressive therapy. We have studied the effect of the ITI of saline (controls) (groups 1 (n = 6) and 3 (n = 6)) or donor splenocytes (groups 2 (n = 10) and 4 (n = 8)) in dogs that received either pharmacological immunosuppression (with cyclosporine and prednisone, +/- azathioprine/cyclophosphamide) (groups 1 and 2) or rabbit anti-dog antithymocyte globulin (groups 3 and 4) at the time of ITI. Kidney or heart allografting (from the donor of the splenocytes) was carried out 16-74 days after ITI; all but four transplants were performed within 16-22 days after ITI. Mean kidney allograft survival was 6, 10, 9, and 9 days, respectively, in groups 1-4. Mean cardiac allograft survival was 7, 14, 8, and 7 days, respectively. There was no statistical difference in allograft survival between those dogs that received ITI of saline and those that received donor splenocytes. These results would suggest that the protocols developed to date using ITI in rodent species may not be successful in dogs.


Subject(s)
Graft Rejection/prevention & control , Graft Survival/immunology , Heart Transplantation/immunology , Kidney Transplantation/immunology , Spleen/transplantation , Thymus Gland/immunology , Animals , Antilymphocyte Serum/biosynthesis , Dogs , Heart Transplantation/pathology , Kidney Transplantation/pathology , Lymphocyte Depletion , Spleen/cytology
15.
Cytometry ; 22(1): 16-21, 1995 Mar 15.
Article in English | MEDLINE | ID: mdl-7587728

ABSTRACT

Induction protocols for pediatric renal transplant recipients commonly utilize polyclonal or monoclonal agents in sequence with additional immunosuppression. Polyclonal agents Minnesota antilymphoblast globulin (MALG), anti-thymocyte globulin (ATGAM) effect both T and B cells while monoclonal agents (OKT3) are T-cell specific. Flow cytometric analysis of T-cell subsets has become the marker of adequacy of immunosuppression during OKT3 therapy, yet to date no marker exists to measure the adequacy of immunosuppression with polyclonal agents. At the University of Michigan, flow cytometric analysis in pediatric renal transplant recipients undergoing polyclonal induction reveals that CD3, CD4, and CD8 suppression initially occurs. As opposed to OKT3, a rebound of CD3 cells occurs despite daily use of a polyclonal agent in sequence with additional immunosuppressives. During these analyses, a single kidney was lost which flow cytometry failed to predict. No significant difference in flow cytometric patterns occurred when comparing ATGAM to MALG induction. Flow cytometry utilized during polyclonal induction in pediatric renal transplant recipients revealed a variety of T-cell suppression patterns but failed to demonstrate persistence of suppression. Despite this lack of suppression as indicated by flow cytometry, a 97% 1 year allograft survival rate exists in the pediatric transplant program at the University of Michigan Medical Center. Therefore, the role of flow cytometry during polyclonal induction has yet to be well defined.


Subject(s)
Antilymphocyte Serum/biosynthesis , Antilymphocyte Serum/therapeutic use , Flow Cytometry , Kidney Transplantation/immunology , Antibody Formation , Child , Evaluation Studies as Topic , Graft Rejection/prevention & control , Humans , Immunosuppression Therapy , Retrospective Studies
16.
Biull Eksp Biol Med ; 116(7): 61-3, 1993 Jul.
Article in Russian | MEDLINE | ID: mdl-8400187

ABSTRACT

Apheresis was applied in 17 patients with hypoplastic anemia, myelodysplastic syndrome, chronic renal failure and other diseases sensitized to HLA. Apheresis was done for removal of anti-HLA antibodies and prevention of nonhemolytic transfusion reactions. Multiple massive apheresis led to a marked decrease in the antibody level. After the first or second apheresis with removal of 700.0-2000.0 ml of plasma weekly half of the patients showed increasing titres of lymphocytotoxic antibodies.


Subject(s)
Antilymphocyte Serum/biosynthesis , Isoantibodies/biosynthesis , Plasmapheresis , Antilymphocyte Serum/blood , Female , HLA Antigens/immunology , Hematologic Diseases/immunology , Hematologic Diseases/therapy , Humans , Isoantibodies/blood , Male
17.
J Autoimmun ; 6(1): 121-30, 1993 Feb.
Article in English | MEDLINE | ID: mdl-8457284

ABSTRACT

Experimental and spontaneous autoimmune disease in animals can effectively be prevented and treated by application of pathogenic autoreactive T cells in an attenuated form. This approach has become known as T cell vaccination. T cell vaccination exploits specifically the ability of the immune system to regulate its autoreactive T cells by mechanisms of network control. The success of T cell vaccination in a variety of rodent animal models has raised hopes for its use as an effective and specific therapy in human autoimmune disease. The aim of this study was to induce an anti-T cell response by T cell vaccination in humans and primates as a pre-clinical study into the feasibility and toxicity of T cell vaccination. Using bulk cultures of T cells from the peripheral blood or an inflamed joint, it was possible to induce a T cell response specific for the injected vaccine and its activation state both in rhesus monkeys and in two patients with active rheumatoid arthritis. In one of the patients there was already a spontaneous T cell response against a mitogen driven T cell line from the peripheral blood, but not against a control T cell line specific for tetanus toxoid, suggesting that regulatory T cell networks are operative in patients with autoimmune disease. Significant clinical effects or side-effects were not observed. The results suggest that T cell vaccination in humans is feasible and non-toxic. It is likely to influence an already ongoing regulatory process. Conditions for making T cell vaccination an effective therapy need still to be worked out by further studies both in primates and in less complex human immune processes.


Subject(s)
Arthritis, Rheumatoid/therapy , Autoimmune Diseases/therapy , T-Lymphocytes/immunology , Vaccination , Animals , Antilymphocyte Serum/biosynthesis , Arthritis, Rheumatoid/prevention & control , Feasibility Studies , Female , Humans , Lymphocyte Cooperation , Macaca mulatta/immunology , Male , Middle Aged
19.
Vet Immunol Immunopathol ; 33(1-2): 163-9, 1992 Jun.
Article in English | MEDLINE | ID: mdl-1632077

ABSTRACT

Complement fixing antibodies were measured in sera from animals immunised with either Theileria annulata sporozoites or autologous or allogeneic schizont-infected mononuclear cells using a complement-mediated micro-cytotoxicity test. The test demonstrated the presence of anti-lymphocyte antibodies in allogeneic cell-immunised animals, which were not detectable in autologous cell- or sporozoite-immunised animals; also that these antibodies were directed to T. annulata-infected and (MHC) class I antigens. Their potential importance in repeated immunisations is discussed.


Subject(s)
Antilymphocyte Serum/biosynthesis , Immunization , Theileria annulata/immunology , Theileriasis/immunology , Animals , Antibody Specificity , Antilymphocyte Serum/analysis , Cattle , Leukocytes, Mononuclear/immunology , Leukocytes, Mononuclear/parasitology , Theileria annulata/growth & development , Theileriasis/blood , Theileriasis/parasitology
20.
J Clin Invest ; 84(3): 907-14, 1989 Sep.
Article in English | MEDLINE | ID: mdl-2474577

ABSTRACT

Hybridization of murine myeloma cells P3-X63-Ag8.653 with splenocytes from a BALB/c mouse immunized with the syngeneic anti HLA-DR1,4,w6,w8,w9 MAb AC1.59 resulted in the development of 108 hybridomas secreting antiidiotypic antibodies. 100 of them inhibited the binding of MAb AC1.59 to target cells. Detailed analysis of the antiidiotypic MAb F5-444, F5-830, F5-963, F5-1126, F5-1336, and F5-1419 showed that all of them recognize idiotopes within or spatially close to the antigen combining site of MAb AC1.59. In cross-blocking experiments, the six antiidiotypic MAbs cross-blocked each other. It is likely that the six MAbs recognize spatially close, but not identical idiotopes because they elicited antiantiidiotypic antibodies of different or similar, but not identical specificity and differ in their ability to elicit anti-HLA class II antibodies. The latter, which were found only in sera from BALB/c mice immunized with antiidiotypic MAb F5-444 and F5-830, mimic the specificity of MAb AC1.59 and express the idiotope defined by the immunizing antiidiotypic MAb. These results indicate that the MAb F5-444 and F5-830 are antiidiotypes beta and the remaining four are antiidiotypes gamma.


Subject(s)
Antibodies, Anti-Idiotypic/analysis , Antibodies, Monoclonal/analysis , Epitopes/analysis , HLA-DR Antigens/analysis , Immunoglobulin Idiotypes/immunology , Animals , Antibodies, Anti-Idiotypic/biosynthesis , Antibodies, Anti-Idiotypic/physiology , Antibodies, Monoclonal/biosynthesis , Antibodies, Monoclonal/physiology , Antilymphocyte Serum/analysis , Antilymphocyte Serum/biosynthesis , Antilymphocyte Serum/physiology , Binding Sites, Antibody , Binding, Competitive , Epitopes/immunology , HLA-DR Antigens/immunology , Hybridomas/analysis , Immune Sera/analysis , Immunoglobulin Variable Region/analysis , Mice , Mice, Inbred BALB C
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