Jerome H. Herman

In vitro studies in reproductive immunology. 1. Suppression of cell-mediated immune response by human spermatozoa and fractions isolated from human seminal plasma.

Abstract Components of the male mammalian genital tract are capable of inhibiting in vitro cell-mediated immune responses. In the current study, intact human spermatozoa in a concentration of 10 7 cells/ml inhibited both spontaneous and PHA-M- and Con A-induced blast transformation of normal human peripheral blood lymphocytes. Similar inhibitory properties were found in human seminal plasma using concentrations of 100 and 400 μg/ml. Sephadex G-100 chromatography of seminal plasma yielded five fractions (F 1 - F 5 ) which were qualitatively assessed by SDS-polyacrylamide disc gel electrophoresis. Fractions 1 and 4 inhibited spontaneous blast transformation while fractions 1 and 3 inhibited mitogen-induced blastogenesis. Normal leukocyte migration was not affected by fractions. It is concluded that suppression of blast transformation might be due to a spermatozoal or seminal plasma component(s) able to bind receptors on lymphocytes specific for PHA-M and Con-A.

Immunopathologic Studies in Relapsing Polychondritis

Serial studies have been performed on three patients with relapsing polychondritis in an attempt to define a potential immunopathologic role for degradation constituents of cartilage in the causation and/or perpetuation of the inflammation observed. Crude proteoglycan preparations derived by disruptive and differential centrifugation techniques from human costal cartilage, intact chondrocytes grown as monolayers, their homogenates and products of synthesis provided antigenic material for investigation. Circulating antibody to such antigens could not be detected by immunodiffusion, hemagglutination, immunofluorescence or complement mediated chondrocyte cytotoxicity as assessed by (51)Cr release. Similarly, radiolabeled incorporation studies attempting to detect de novo synthesis of such antibody by circulating peripheral blood lymphocytes as assessed by radioimmunodiffusion, immune absorption to neuraminidase treated and untreated chondrocytes and immune coprecipitation were negative. Delayed hypersensitivity to cartilage constituents was studied by peripheral lymphocyte transformation employing [(3)H]thymidine incorporation and the release of macrophage aggregation factor. Positive results were obtained which correlated with periods of overt disease activity. Similar results were observed in patients with classical rheumatoid arthritis manifesting destructive articular changes. This study suggests that cartilage antigenic components may facilitate perpetuation of cartilage inflammation by cellular immune mechanisms.

In vitro studies in reproductive immunology — 2 demonstration of the inhibitory effect of male genital tract constituents on PHA-stimulated mitogenesis and E-rosette formation of human lymphocytes

The effect of male genital tract components (human spermatozoa, intact and chromatographed seminal plasma fractions) on in vitro cell-mediated immune reactions was examined. Their addition to PHA-containing lymphocyte cultures resulted in a marked degree of inhibition of DNA synthesis, the response varying with the fractions employed. The interference by such components with a terminal cell event during blast transformation was suggested by the failure of stimulated cultures to incorporate thymidine irrespective of the time during the incubation period at which the constituents were added. Seminal plasma and sperm extracts were also shown to inhibit T-cell associated E-rosette formation. The inhibitory properties of genital fractions remained unaltered after repetitive freeze-thaw cycling when tested in the transformation and E-rosette assays. Heating to 100 degrees C modified the response with some of the fractions.

The Effect of Human Spermatozoa on Antigen and Mitogen Induced Blastogenesis

In an attempt to identify factors capable of specifically or nonspecifically modulating results in an in vitro blastogenic assay system, studies were performed evaluating cell mediated immune response to human spermatozoa. Spermatozoa were capable, on a dose dependent basis, of both inhibiting and stimulating normal lymphocyte DNA synthesis as well as suppressing mitogen-induced response. Pretreatment of spermatozoa with neuraminidase and alpha-methyl-D-mannoside abrogated suppressive properties on spontaneous and/or mitogen-induced stimulation. Such suppressive activity on mitogen-induced response was observed using B and T-cell enriched populations. Inhibitory properties were not evident using intact cell populations stimulated by specific antigen. It is suggested that: (1) receptors on spermatozoa bind lectin; and (2) such receptors can be inactivated by enzymes having glycoprotein specificity.

Immunobiology of cartilage

Abstract Although conflicting data have been presented, several definitive conclusions may be reached. Success may be anticipated using intact autogenous cartilage grafts of varying structure if septic and mechanical problems can be circumvented. In contrast, the allogeneic grafting of such tissue has a high failure rate, even in conjunction with host immunosuppression. Chondrocytes contain major histocompatibility antigens, as well as perhaps surface determinants unique to their cell type. Though an immune response may be generated against such antigens, matrix is able to afford protection, the degree dependent upon prior host sensitization to donor tissue. Thus cartilage appears to be an “immunologically privileged tissue”, not because of its lack of antigenicity but because of the sequestration of its antigens from the immune system as reflected by matrix protection of chondrocytes and the structural conformation of matrix molecules. It is evident that better designed immunologic investigations, including studies pertaining to matrix antigenicity as well as more directly to chondrocyte surface determinants, are indicated in this important area.

Nonsteroidal Anti-Inflammatory Drugs and Modulation of Cartilaginous Changes in Osteoarthritis and Rheumatoid Arthritis Clinical Implications

Nonsteroidal anti-inflammatory drugs have a potential for modifying the complex pathophysiologic events leading to cartilage destruction in various forms of arthritis. Following an evaluation of basic mechanisms in the pathogenesis of cartilaginous destructive lesions, the effects of nonsteroidal anti-inflammatory drugs on normal chondrocyte metabolism are discussed. Their capacity to modulate cartilage and bone lesions in experimental forms of arthritis is addressed, as is the manner in which they may modify the pathophysiology of cartilage destruction in human forms of arthritis. Different classes of nonsteroidal anti-inflammatory drugs produce different effects in certain in vivo or in vitro settings.

Modulation of human lymphocyte response by cartilage proteoglycans and glycosaminoglycans—A comparison between normal subjects and patients with rheumatoid arthritis☆

Abstract With recognition of the effectual capacity of microenvironment in influencing the inductive and effector expression of immunologic function, studies have been undertaken to determine the potential regulatory role of basic connective tissue constituents on spontaneous and phytomitogen-induced lymphocyte DNA synthesis. Proteoglycan constituent fractions and glycosaminoglycans derived from normal human cartilage were shown capable of modulating spontaneous DNA synthesis and the PHA responsiveness of peripheral blood lymphocytes derived from normal subjects and patients having destructive rheumatoid arthritis. Dependent upon the fraction employed, its concentration, and duration of cell exposure, spontaneous as well as mitogen-induced synthesis could be stimulated and/or suppressed without significant effect upon lymphocyte viability. Under conditions in which connective tissue fractions themselves induced suppressive effects on synthesis, the affected population was more responsive in a relative or absolute manner to mitogenic stimulation. Reciprocally, direct lymphocyte stimulation was associated with suppressed mitogen responsiveness. Rheumatoid response clearly differed from that of normal subjects.

Cytokine modulation of chondrocyte metabolism--in vivo and in vitro effects of piroxicam.

Changes in the macromolecuaar proteolycan (PG) and collagen of the cartilage matrix may culminate in irreparable tissue destruction. Molecular modifications appear to result from: (A) exogenous proteinases, (B) endogenous chondrocyte proteinases whose synthesis and release is modulated by exogenous non-enzymatic cytokines (CKs) and (C) quantitative and/or qualitative alterations in chondrocyte PO and collagen synthesis which are potentially induced by exogenous CKs. Studies have recently been initiated to determine the effect of piroxicam on the synthesis and activity of such metabolic regulatory CKs in patients with rheumatoid arthritis and osteoarthritis, and in age-, sex-, and racematched controls. Therapeutic doses of piroxicam alone had no effect on the anabolic or cataboiic function of chondrocytes. Current studies concern the effect of piroxicam on: (a) spontaneous and lectin-dnven production by peripheral blood monocytes and T-cells of trypsin-sensitive, heat-labile CKs (interleukin 1, lymphokine) which, in a protein-and RNA-synthesis-dependent manner, induce a concentration and duration of substrate exposure dependent release of chondrocyte PG-and collagendegrading neutral proteinases in cartilage organ and chondrocyte suspension culture systems; (b) spontaneous and lectin-driven synthesis by peripheral blood T-cells of lymphokines capable of suppressing chondrocyte PG, glycosaminoglycan, protein, collagen and nucleic acid synthesis in a quantitatively reversible manner; (c) pathological synovial membrane synthesis of such cataboiic-inducing and anabolic-modulatory CKs. These experimental model systems are reviewed together with preliminary data on the effect of piroxicam.

Inhibition of mitogen induced blast transformation by male genital components.

Publisher Summary This chapter discusses the inhibition of mitogen-induced blast transformation by male genital components. Human and guinea pig seminal plasma can cause specific and nonspecific inhibition of in vitro cell-mediated immune (CMI) responses. Following experimental or natural sensitization to spermatozoa, a specific antibody and a CMI response occurs. In a study described in the chapter, human peripheral blood lymphocytes were isolated from whole heparinized blood, drawn from normal donors, using Ficoll–Hypaque gradient centrifugation. The cells obtained were used for the microblast transformation and leukocyte inhibition tests procedures. Polyhydroxyalkanoates (PHA)- and spontaneous-induced blast transformation were found to be significantly inhibited by 100 and 400 μg/ml human seminal plasma. The degree of the inhibition of PHA and concanavalin A-induced blast transformation by human spermatozoa is decreased by saturating the spermatozoa active sites with higher amounts of mitogens. Male genital components are immunosuppressive if used in the in vitro CMI systems. These components have antigenic specificity and concurrent suppressive properties, which can mitigate against documentation of immune responses to them.

IMMUNOLOGICAL CONSEQUENCES OF HUMAN VASECTOMY

A 2-year followup study of 41 men undergoing vasectomy was initiated at the University of Cincinnati Medical Center Cincinnati Ohio in 1972 in an attempt to evaluate the change in the humoral and cellular immunological systems which might result from potential sustained genital antigen exposure induced by vasectomy. These 41 subjects - Group 1 - have now been followed for up to 4 years. The 6 major components to the study included complete medical evaluation: tests performed pre-vasectomy and at 6 weeks 3 6 12 and 24 months post-vasectomy regarding humoral factors; immune complexes; genital antibodies; cellular immune system; and coagulation studies. It became clear that changes in many of the immunological parameters could be related to a multitude of exogenous factors occurring during the 2-year period making it difficult to interpret data which compared 1 pre-vasectomy observation with 5 post-vasectomy observations. To deal with this a 2nd study group in which 18 vasectomized subjects would be matched with a non-vasectomized age and socioeconomic control group was initiated in 1975. Results of the studies on the 36 group 2 subjects included the following: 1) there were no abnormalities in any of the subjects at 12 months post-vasectomy; 2) regarding humoral factors 1 vasectomized subject has a positive latex test 6 weeks post-vasectomy and a 2nd subject had thyroid and gastric parietal cell antibodies post-vasectomy and the other 34 subjects showed no abnormalities; 3) tests for immune complexes were negative in both groups; 4) 60% of the vasectomized group became positive in the Kibrick sperm agglutination test post-vasectomy; only 1 control subject became positive; 5) skin test responses in control and vasectomized groups were similar and the blastogenic lymphocyte responses to mitogens showed variability in both groups. No clinical evidence of autoimmune or other specific disorder has appeared during the followup nor have there been specific abnormalities of humoral or cellular systems.