Production Of Blastogenic Factor Research Articles

Blastogenic factor production by guinea pig B lymphocytes stimulated with thymus-dependent antigen.

The purpose of the present paper is to determine whether antigen-stimulated B lymphocytes could produce blastogenic factor (BF) in the system of guinea pigs (strain 2) sensitized with keyhole limpet hemocyanin (KLH) or ovalbumin (OVA), known as thymus-dependent antigen, and, further, whether macrophages (M phi) are required for antigen activation of B cells to produce this lymphokine. Additionally, the effect of BF on normal T and B cells was evaluated. B lymphocytes were purified by double nylon wool adherence, plastic dish adherence. Sephadex G-10 column passage and sedimentation of E rosette forming cells on Ficoll-Isopaque. KLH- or OVA-primed B lymphocytes as well as T lymphocytes could produce BF when stimulated with specific antigen in vitro, and M phi were required for antigen activation of both T and B cells to produce BF. The optimal BF production was observed at a T or B cells: M phi ratio of 1:0.1 (in the system of KLH sensitization) or 1:0.5 (in OVA sensitization). BF production by B cells appeared to be less than that by T cells. BF produced by B cells was able to stimulate both normal T and B cells to proliferate; the proliferative responses of normal B cells to both B-cell-derived BF and T-cell-derived BF were significantly greater than those of normal T cells to these BFs. Readdition of M phi was not required for the T or B cell response to BF.

Suppression of experimental allergic orchitis and cellular immune response in the guinea pig by pretreatment with testis antigen in incomplete Freund's adjuvant.

Pretreatment of guinea pigs with homologous testicular antigen (TA) in incomplete Freund's adjuvant (IFA) markedly reduced both the incidence and severity of experimental allergic orchitis (EAO) when animals were subsequently challenged with TA in complete Freund's adjuvant (CFA) and examined at 2 weeks after challenge. The pretreatment rendered lymph node lymphocytes, particularly T cells, specifically unresponsive to the subsequent orchitogenic challenge as judged by antigen-induced in vitro lymphocyte proliferative response and blastogenic factor production, whereas antisperm antibody formation was not affected. The suppression induced was of a transitory nature, lasting 3 weeks after orchitogenic challenge, but between 4 and 5 weeks after challenge the development of EAO and aspermatogenesis became evident in parallel with the restoration of cellular immune responsiveness. Cyclophosphamide treatment 3 days before orchitogenic challenge abolished the preventive effect of pretreatment with TA in IFA. Transfer of serum obtained from pretreated animals failed to prevent actively sensitized recipients from developing EAO, which does not favor a possible occurrence of serum blocking factors in protected animals.

T cell recognition of Moloney sarcoma virus proteins during tumor regression. I. Lack of a requirement for macrophages and the role of blastogenic factors in T cell proliferation.

Abstract The characteristics of the T cell blastogenic response of lymphocytes from Moloney leukemia/sarcoma virus inoculated mice against the virion proteins gp71 and p12 were examined. The proliferative responses to both antigens were independent of a requirement for macrophages as determined by macrophage-depletion experiments, the lack of an effect of antisera against Ia on proliferation and by analysis of the number of cell interactions required for proliferation. The kinetics of antigen-lymphocyte interaction were examined and found to require only 4 to 6 hr for half-maximal activation. Proliferation in response to gp71 or p12 was found to be due to the production of blastogenic factor(s), and using additivity experiments, little if any proliferation appears to be associated with antigen-specific lymphocytes. The production of blastogenic factor(s) does not require proliferation and is temporally displaced from antigen-lymphocyte interaction by 8 to 10 hr.

Development of Cell-Mediated Hypersensitivity in Guinea Pigs following Infection with <i>Nocardia asteroide</i><i>s</i>

Partially purified protein antigen which was prepared from cells of <i>Nocardia asteroides</i> was used to assess the immune responsiveness of guinea pigs inoculated subcutaneously with living <i>N. asteroides</i>. Cell-mediated reactivity, as measured by in vitro lymphocyte transformation of and blastogenic factor production by lymph node cells as well as a delayed skin reaction, appeared clearly at 1 week after <i>N. asteroides</i> infection, when an early abscess was formed at the inoculated site. The in vitro proliferative response of lymph node cells reached a maximum at 2 weeks at which time the lesion of the inoculated site showed central suppuration surrounded by epithelioid and giant cells. The lymphocyte transformation response then declined rapidly but a significant response continued until 6 weeks after infection. The production of blastogenic factor was maximum at 3 weeks, but continued till 6 weeks. A high level of skin reactivity was maintained until 6 weeks. By 6 weeks after infection the lesion at the inoculated site usually resolved and disappeared. Thus, enhancement of cell-mediated reactivity coincided with the time of granulomatous conversion of an early suppurative abscess at the site of inoculation of <i>Nocardia</i> organisms. In studies on the lymphocyte proliferative response in vitro of <i>N. asteroides</i>-infected animals, purified lymph node T cell populations failed to respond to <i>Nocardia</i> antigen, but they could be induced to proliferate in the presence of autologous macrophages or B cells. Purified B lymphocytes, though unresponsive to <i>Nocardia</i> antigen by themselves or following the addition of macrophages, were stimulated by <i>Nocardia</i> antigen in the presence of T cells. The proliferative response of lymph node cells to <i>Nocardia</i> antigen seems to be the result of a collaborative event that occurs between T cells and macrophages or T and B cells.

Effect of Treponema pallidum-infected testis supernatants on the cellular response of normal rabbit lymphocytes.

This report presents evidence indicating that both Treponema pallidum and the T. pallidum-free supernatant from T. pallidum-infected rabbit testes exert an immunosuppressive effect on the spontaneous proliferation and mitogenic response of normal rabbit lymphocytes. This effect seemed to be on the level of clonal expansion since antigenic recognition and blastogenic factor production occurred in spite of the presence of the immunosuppressive substance(s). The implication of this finding for T. pallidum multiplication in rabbit testes is discussed.

Macrophage Requirements of PPD‐induced Blastogenic Factor Production by T and B Lymphocytes in the Guinea‐Pig

An attempt has been made to characterize the cell type which releases blastogenic factor(s) (BF) on purified protein derivative (PPD) stimulation in complete Freund's adjuvant (CFA)-sensitized guinea-pig system and, further, to examine whether macrophages (Mphi) are required for antigen activation of T or B lymphocytes to produce this lymphokine. Both T and B lymphocyte populations from guinea-pigs 2-3 weeks after CFA sensitization (early-stage T and B cells) failed to produce significant amounts of BF in response to PPD, whereas both the populations from 10 to 14 weeks after sensitization (late-stage T and B cells) acquired the ability to produce BF by themselves. It is noteworthy that readded autologous Mphi not only functioned well to augment BF production by the late-stage T and B cells but also completely made up for the inability of the early-stage T and B cells to produce BF. From these data it is reasonable to conclude that in the guinea-pig sensitized T and B cells can both release BF on exposure to PPD and that Mphi either reconstitute or augment the BF synthetic response of T and B cells.

DECREASED BLAST CELL TRANSFORMATION AND BLASTOGENIC FACTOR PRODUCTION IN CORD BLOOD LYMPHOCYTES

Increased susceptibility of newborn to infection with agents controlled by cell-mediated immunity (CMI) is well documented. To evaluate the CMI function of newborn we studied the peripheral T cell numbers by E-rosette technique, as well as the phytohemagglutinin (PHA) lymphocyte transformation and the production of blastogenic factor in the cord blood from 11 healthy newborn infants and in venous peripheral blood from 6 healthy adults. It was found that 1. T cell numbers in newborn were comparable to the ones found in the adult controls (mean 50% in newborns and 52% in adults); 2. the PHA lymphocyte transformation as measured by tritiated thymidine incorporation was markedly lower than in adults (mean PHA index 16 In the cord blood and 70 in adults), although spontaneous transformation was much higher in newborn compared to adult lymphocytes; and 3. the supernatant derived from PHA stimulated cord lymphocytes (after removal of PHA) failed to stimulate neonatal lymphocytes as measured by tritiated thymidine incorporation (mean blastogenic index 1.6, range 1-2) in contrast to the supernatant of PHA stimulated adult lymphocytes which significantly stimulated the neonatal lymphocytes (mean blastogenic index 5.1, range 3.5-7). These data suggest that newborn lymphocytes, while possessing the surface properties of T cells, are functionally immature.

Dissociation of blastogenic factor production and lymphocyte transformation of guinea pig lymphocytes stimulated with tuberculin PPD.

In vitro lymphocyte transformation of guinea pig lymph node cells induced by tuberculin PPD was essentially eliminated by treatment with 5-bromo-2-deoxyuridine and light. The remaining cell population was still capable of producing blastogenic factor and further, the amount of blastogenic factor released was comparable to that from the intact population in the same interval of culture. This result favors the conception that cells producing blastogenic factor in response to antigen are distinct from the cells responding to the antigen by DNA synthesis and proliferation.

MLC stimulatory capacity and production of blastogenic factor in patients with chronic lymphatic leukemia and Hodgkin's disease

The aim of the present study is to evaluate the stimulatory capacity of chronic lymphatic leukemia (CLL) and Hodgkin's disease (HD) lymphocytes in the one-way mixed lymphocyte reaction (MLR) and to investigate whether, in patients with these diseases, a correlation exists between the degree of stimulatory capacity and production of a blastogenic factor. These studies might help to resolve the controversy about the nature of the stimulating cells and to offer some insight into the mechanism of the stimulation in the MLR. The significant observations from this study are summarized as follows: (1) Stimulatory capacity of CLL lymphocytes was intact or increased, while their responding capacity was markedly depressed. (2) In our group of patients with advanced Hodgkin's disease, both stimulating and responding capacities were imparied. (3) In these patients a correlation existed between the degree of stimulatory capacity and production of a blastogenic factor. This observation would raise the possibility that the production of a blostogenic factor may in some way be involved in the stimulation of responding lymphocytes in mixed cultures. Our results, which show a dichotomy between MLC stimulatory and responding capacities of CLL lymphocyte, may suggest that different factors are involved in stimulation and response.

Production of blastogenic factor by mitomycin-treated cells in mixed lymphocyte cultures.

Production of blastogenic factor by mitomycin-treated cells in mixed lymphocyte cultures.