Analysis of B lymphocyte function in prospective renal transplant patients

Degiannis, Dimitrios (1986) Analysis of B lymphocyte function in prospective renal transplant patients. PhD thesis, University of Glasgow.

Abstract

It is well established that uraemia impairs the in vivo cell-mediated immune responses while the in vitro lymphocyte responses of patients on maintenance dialysis have been reported to be impaired or normal . The work in this thesis deals predominantly with production of immunoglobulin in vitro by peripheral blood mononuclear cells obtained From normal and uraemic subjects. The protein A plaque Forming cell assay was utilised to enumerate immunoglobulin secreting cells and several parameters affecting its performance were examined. Thus, it was Found that plaque Forming cell numbers were dependent on the concentrations of sheep red blood cells, anti-serum, DEAE-dextran and complement while PEG 6000 was shown to improve the quality of the plaques. The optimal conditions For storing the cells before plating and incubating them after plating were also established and in addition, it was shown that cryopreservation does not affect plaque Forming cel1 activity. The optimal conditions For both proliferation and immunoglobulin production in response to mitogenic stimulation were also examined and once again the suitability of cryopreserved peripheral blood mononuclears For use in mitogen-stimulated cultures was confirmed, A normal range For both spontaneous and mitogen induced plaque Farming cells was established and it was shown that pokeweed mitogen(PWM) was the most effective polyclonal B cell activator in inducing plaque Forming cells while Epstein-Barr-Virus(EBU) induced similar IgM-PFC as PWM. Co-stimulation of control peripheral blood mononuclear cells with Concavalin A(ConA) showed that both PWM and Staphylococcus Aureus Cowan I(SAC) cultures were suppressed although the Former proved to be much more sensitive. Steroids are known to inhibit suppressor T cells and when methyl-prednisolone was added in PWM and SAC cultures of control peripheral blood mononuclear cells, PWM induced production of immunoglobulin was more readily enhanced. I propose that these polyclonal B cell activators differ in their susceptibility to regulation by suppressor T cells. When indomethacin was added in the cultures to inhibit prostaglandin synthesis, it was observed that SAC responses were slighty but non-significantly enhanced. Uraemia was not Found to have a significant effect on the number or proportion of lymphocyte populations but . it impaired the proliferative' responses to PHA and PUIM. However, the relative in vitro immunosuppressive effect of steroids was not Found to be stronger in these cultures. EBU and PWM induced plaque Formation was normal in uraemic patients while both spontaneous and SAC induced immunoglobulin production were reduced. In addition, the combination of PWM and SAC did not produce the synergistic effect Found in control cultures. Pre-incubation of control cells with uraemic serum affected the quality and suppressed slightly the numbers of PWM-induced plaque Forming cells. Addition of methyl-prednisolone to SAC cultures did not produce the enhancing effect Found in controls indicating that the reduced SAC responses in uraemia were not caused by incraasad supprassar T cell activity. Howevar, the SAC responses of uraemic peripheral blood mononuclear cells seemed to benefit more by the addition of indomethacin than the SAC responses of control cells. My results indicate strongly that the reduced production of immunaglobulin in uraemia is not due to reduced numbers of lymphocytes but reflects a true functional abnormality and the observed reduced proliferative responses of uraemic peripheral blood mononuclears to T cell mitogens indicate that T lymphocyte function is impaired. The studies using polyclonal B cell activators suggested that B cell function and T-B cell co-operation remained intact in uraemic patients and indicated that the defective immunoglobulin responses in both the spontaneous and SAC system were due to a functional abnormality of the helper T cell.

• Item Type: Thesis (PhD)
• Qualification Level: Doctoral
• Keywords: Immunology.
• Colleges/Schools: College of Medical Veterinary and Life Sciences
• Supervisor's Name: Mowat, Dr. Allan
• Date of Award: 1986
• Depositing User: Enlighten Team
• Unique ID: glathesis:1986-72070
• Copyright: Copyright of this thesis is held by the author.
• Date Deposited: 24 May 2019 15:11
• Last Modified: 15 Jun 2021 10:19
• URI: https://theses.gla.ac.uk/id/eprint/72070