Association of certain Ia allotypes with the occurrence of chronic lymphocytic leukemia. Recognition by a monoclonal anti-Ia reagent of a susceptibility determinant not in the DR series.

The Ia antigen allospecificities of individuals with B type chronic lymphocytic leukemia differed significantly from those of a control population. A monoclonal antibody, IVD12, directed to a MB3-1ike determinant, reacted with 92.5 percent of the individuals with leukemia and yielded the greatest positive relative risk, 13.5. A lower degree of positive association was found with the presence of the MT2 determinant. In contrast, the low observed frequency of the MT1/MB1 determinant among leukemic individuals was associated with the most significant negative relative risk, -8.1. Among HLA- DR specificities, the relative risk associated with the presence of DR5 was positive while that with DR2 was negative.

Evidence for the separate molecular expression of four distinct polymorphic Ia epitopes on cells of DR4 homozygous individuals.

The monoclonal antibodies 109d6 and IVD12 reacted with separate polymorphic Ia epitopes in immunofluorescent studies. Using a panel of lymphoblastoid B cell lines, antibody 109d6 reacted with all HLA-DR4 or DR7 positive lines in a pattern resembling the MT3 specificity recognized by human alloantisera. The antibody IVD12 reacted with all HLA-DR4 and two of three DR5 positive B cell lines suggesting that it recognized a specificity similar to MB3. The intensity of fluorescence was greater on DR5(+) cell lines than on DR4(+) cell lines relative to the amount of a nonpolymorphic Ia determinant. Among 45 unrelated control individuals reactivity with antibody 109d6 was correlated most closely with (r = 0.724) but not identical in occurrence to the MT3 specificity. Cocapping experiments demonstrated that the 109d6 epitope and the IVD12 epitope were present on independently redistributed cell surface molecules of DR4 homozygous lymphoblastoid cell lines. Furthermore, the DR4 alloantigens detected by an absorbed polyclonal human alloserum were similarly identified on molecules that were independent from those bearing either the 109d6 epitope or the IVD12 epitope. Taken together, these data indicate the existence of at least four distinct, serologically defined Ia molecular species.

The cellular basis of impaired T lymphocyte functions in the elderly.

Immunologic changes associated with aging were studied by various immunologic tests in 24 aged persons (age range, 76-83) and 25 young persons (age range, 20-40). The responses to phytohemagglutinin (PHA) and concanavalin A (Con A) were depressed in the aged subjects compared to the young ones (p less than 0.05), whereas the responses to pokeweed mitogen (PWM) were similar. The activity of adhereent and non-adherent cells was assessed in various combinations. The adherent cells of aged persons were indistinguishable from those of young persons in their ability to response to Con A. Lymphocytes from the aged synthesized larger in vitro amounts of immunoglobulin than did lymphocytes from the young, when stimulated with PWM. Con A-stimulated T lymphocytes derived from aged subjects showed a variable loss of suppressor activity. The mixed lymphocyte culture reaction with mitomycin-treated allogeneic and autologous cells was also impaired in aged subjects. Such an impaired response in the aged is related to higher incidences of malignant lesions and auto-antibodies.

Enhancement of lymphocyte response to PHA by lysosomal enzymes from polymorphonuclear leukocytes of RA joint fluid. I. Biological effect on T lymphocyte function.

The effect of polymorphonuclear leukocyte (PMN) granule lysates obtained from joint fluid of RA on the in vitro DNA synthesis of PHA-stimulated autologous lymphocytes from joint fluid was studied. Lymphocytes were cultured for 3 days with or without PMN lysates in 2 ml of RPMI-1640 supplemented with 10% heat-inactivated fetal calf serum (FCS). The lymphocytes were stimulated with phytohemagglutinin (PHA-M). The DNA synthesis was measured by counting the [3H]thymidine incorporation. Lymphocytes from RA joint fluid stimulated with PHA-M showed 19,466+/-987 cpm (mean+/-SE) per 10(6) cells in the absence of PMN lysates. Upon addition PMN lysates to the PHA-stimulated lymphocytes, the maximum in vitro DNA synthesis increased to 44,877+/-1338 cpm. The enhancing effect of PMN lysates was abolished by plasma inhibitors or by passage through a column of protease inhibitor (Trasylol). It was concluded, therefore, that the enhancing effect of PMN lysates on PHA-stimulated lymphocytes may be associated with lysosomal proteases. Based on experiments using separated T and B lymphocytes, the enhancing effect of PMN lysates was considered to result from the activation of T lymphocytes. The results obtained in the present study suggest an important role for lysosomal proteases in the perpetuation of rheumatoid synovitis.

Characteristic and functional specificity of anti-human BAT (brain associated thymocyte antigen) serum.

A rabbit antiserum to human fetal brain after multiple absorption reacted with 100% of thymocytes, 55% of peripheral blood lymphocytes and 90% of enriched T lymphocytes, but not significantly with B lymphocytes. Spontaneous SRBC rosette formation was inhibited by anti-BAT pretreatment, but EAC-rosette formation remained unaffected. The antiserum was itself highly stimulatory. However, cells treated with the antiserum and complement exhibited marked inhibition of responsiveness to Con A, little effect with PHA and no alteration with PWM. The MLC reaction was inhibited only when the responder cells were treated with the antiserum and complement. Treatment of sensitized lymphocytes with the antiserum and complement caused a dose-dependent suppression of blastogenic response to both PPD and n-DNA. No effect, however, was noted in MIF producing cells. Con A induced suppressor function of lymphocytes was abolished by treatment with the antiserum and complement. These results indicate that the anti-BAT serum obtained by us can be utilized for the isolation of T lymphocyte subsets.

Evidence of aberration of T-cell subsets in aged individuals.

In the present study, T-cell subsets from aged individuals were examined by using anti-BAT (brain-associated thymocyte antigen) serum. Anti-BAT serum was raised against the human fetal brain at 28 weeks of gestation. After absorption wit AB erythrocytes, B-cell lines, and leukaemic cells, anti-BAT serum was T cell-specific but unreactive to normal B cells. The ability of anit-BAT serum-treated lymphocytes from aged individuals to respond to concanavalin A, phytohaemagglutinin, and pokeweed mitogen (PWM) was unaltered even at a high concentration. In PWM-stimulated Ig synthesis, T lymphocytes lacking the anti-BAT serum-reactive T-cell subset enhanced the PWM-stimulated Ig synthesis of autologous B lymphocytes from young individuals. The Con A-induced suppressor function of lymphocytes from aged individuals was not significantly abolished by treatment with anti-BAT serum and complement. In the autologous mixed lymphocyte reaction, the decrease in response was minimal when responder cells from aged individuals was treated with anti-BAT serum even at a high concentration. It is concluded that the T-cell subset with suppressor function is defective in aged individuals.

Mitogenic responses to lipopolysaccharide by B lymphocytes from patients with systemic lupus erythematosus.

Peripheral blood lymphocytes from 43 patients with systemic lupus erythematosus (SLE) and from age- and sex-matched normal controls were cultured with lipopolysaccharide (LPS) to examine the response to the polyclonal B-cell activator. Lymphocytes from active SLE patients incorporated 4840 +/- 471 (mean +/- SE) cpm in response to LPS, whereas lymphocytes from inactive SLE patients incorporated 6906 +/- 897 cpm. In contrast, lymphocytes from normal individuals incorporated 7452 +/- 1126 cpm. Ig synthesis of lymphocytes from active SLE in response to LPS stimulation was also less than that of normal individuals. The helper T-cell function of active SLE, as examined by co-culturing irradiated SLE lymphocytes with unirradiated normal lymphocytes, was normal. These results thus suggested that a defect of B lymphocytes exists in active SLE patients. This B-cell defect and T suppressor cells apparently play an important role in the pathogenesis of SLE.

Studies of anti-lymphocyte antibody of patients with active SLE. I. Cause of loss of suppressor T-lymphocyte function.

Effect of anti-lymphocyte antibody of active systemic lupus erythematosus (SLE) on lymphocyte function was examined. Lymphocytes from normal individuals treated with anti-lymphocyte antibody and complement exhibited marked inhibition of response to concanavalin A (Con A), while the response of lymphocytes to phytohaemagglutinin M (PHA-M) and pokeweed mitogen (PWM) was slightly affected. In mixed lymphocyte culture response, both stimulator and responder cells were insensitive to anti-lymphocyte antibody. Treatment of sensitized lymphocytes with anti-lymphocyte antibody and complement caused a dose-dependent suppression of blastogenic response to purified protein derivatives (PPD). No effect, however, was noted on migration-inhibitory factor (MIF)-producing cells. In PWM-driven Ig synthesis, T lymphocytes lacking the anti-lymphocyte antibody-reactive T-cell subset enhanced PWM-driven Ig synthesis of autologous B lymphocytes. Con-A-induced suppressor function of lymphocytes was abolished by the treatment with anti-lymphocyte antibody and complement. The present study demonstrated that lymphocytes from normal individuals after treatment with anti-lymphocyte antibody and complement showed similar immunological reactivities with lymphocytes from active SLE, indicating that those anti-lymphocyte antibodies could play an important role in defective suppressor cell function.

Studies of anti-lymphocyte antibody in patients with active SLE. II. Effect of anti-lymphocyte antibody on autoreactive cell clones.

The effect of anti-lymphocyte antibodies of active systemic lupus erythematosus (SLE) on the immune regulation of autoantibody production was studied. The present study demonstrated that there were native DNA (nDNA)-sensitized T lymphocytes even in inactive SLE and no or few nDNA-sensitized T lymphocytes in normal individuals, and that in the inactive stages of SLE suppressor T lymphocytes might inhibit the activation of nDNA-sensitized T lymphocytes eliciting the production of anti-DNA antibodies by B lymphocytes. In the active stage of SLE, the anti-lymphocyte antibodies could eliminate the suppressor function of T lymphocytes or a subset of cells capable of either regulating their appearance or differentiating into them, which inhibited such responses. The different suppression of DNA and extractable nuclear antigen (ENA)-stimulated blastogenic response is further discussed.

In vitro immune response of SLE lymphocytes. The mechanism involved in B-cell activation.

Peripheral blood lymphocytes from 26 patients with systemic lupus erythematosus (SLE) and six normal individuals were tested for IgG synthesis in the presence or absence of PWM. Lymphocytes from patients with active SLE synthesized increased amounts of IgG in the absence of PWM and reduced amounts of IgG in the presence of PWM. Serum from patients with active SLE had an enhancing effect on the in vitro IgG synthesis of normal lymphocytes. The IgG or F(ab')2 fractions of SLE serum retained the enhancing effect on in vitro IgG synthesis, and the enhancing activity was absorbed by human spleen cells. As little as 4 h of incubation with SLE serum was needed for the enhancing activity of normal lymphocytes. Treatment of B lymphocytes appeared to be of main importance for an increase in the in vitro IgG synthesis of SLE serum-treated lymphocytes. These results suggest that anti-B-lymphocyte antibodies from patients with active SLE are responsible in part for the hyperactive response of SLE B lymphocytes.

Functional differences of anti-T-cell antibody in patients with systemic lupus erythematosus and ulcerative colitis.

The loss of suppressor T-cell function results in an abundant production of autoantibodies in systemic lupus erythematosus (SLE). As a cause of this suppressor T-cell defect, anti-T-cell antibody seems to be of prime importance. On the other hand, anti-T-cell antibodies can be detected in various other autoimmune diseases, but their functional characteristics have not been determined. In the present study, the functional characteristics of anti-T-cell antibody from a selected subgroup of patients with ulcerative colitis (UC) were compared with those from patients with SLE. Anti-T-cell antibody from the patients with SLE reacted with a T8 subset, resulting in a suppressor defect, whereas anti-T-cell antibody from the UC patients reacted primarily with a T4 subset. Functionally, SLE- T cells failed to proliferate in response to concanavalin A, whereas UC- T cells from UC patients failed to proliferate in response to phytohaemagglutinin. In the Ig synthesis system, both SLE- and UC- T cells increased Ig production of B cells. Since UC+ T cells did not contribute to the generation of Con-A-inducible suppressor activity, we believe that serum from the selected subgroup of patients with UC reacted with the inducer T-cell subset.

Association of susceptibility to certain hematopoietic malignancies with the presence of Ia allodeterminants distinct from the DR series; utility of monoclonal antibody reagents.

The Ia antigen allospecificities of individuals with either B type chronic lymphocytic leukemia or hairy cell leukemia resembled one another and differed significantly from those of a control population. In contrast, the Ia alloantigens of individuals with non-Hodgkin's lymphoma were distinctly different from those of the leukemic group but differed little from the control group. A monoclonal antibody, IVD12, directed to an MB3-like determinant reacted with the greatest proportion of the leukemic individuals and yielded the highest positive relative risk. A lower degree of positive association was found with the presence of the MT2 determinant. In contrast, the low observed frequency of the MT1/MB1 determinant among leukemic individuals was associated with the most significant negative relative risk. The relative risk associated with the presence of DR5 was positive, while among patients with chronic lymphocytic leukemia the relative risk associated with DR2 was negative. Among patients with Hodgkin's Disease the relative risk associated with the presence of DR5 was significantly increased.