Cyclophilin, a specific cytosolic binding protein responsible for the concentration of the immunosuppressant cyclosporin A by lymphoid cells, was purified to homogeneity from bovine thymocytes. Cation-exchange high-performance liquid chromatography resolved a major and minor cyclophilin species that bind cyclosporin A with a dissociation constant of about 2 X 10(-7) moles per liter and specific activities of 77 and 67 micrograms per milligram of protein, respectively. Both cyclophilin species have an apparent molecular weight of 15,000, an isoelectric point of 9.6, and nearly identical amino acid compositions. A portion of the NH2-terminal amino acid sequence of the major species was determined. The cyclosporin A-binding activity of cyclophilin is sulfhydryl dependent, unstable at 56 degrees C and at pH 4 or 9.5, and sensitive to trypsin but not to chymotrypsin digestion. Cyclophilin specifically binds a series of cyclosporin analogs in proportion to their activity in a mixed lymphocyte reaction. Isolation of cyclophilin from the cytosol of thymocytes suggests that the immunosuppressive activity of cyclosporin A is mediated by an intracellular mechanism, not by a membrane-associated mechanism.

https://science.sciencemag.org/content/sci/226/4674/544.full.pdf

Cyclophilin: A Specific Cytosolic Binding Protein for Cyclosporin A

The present invention is directed to secreted and transmembrane polypeptides and to nucleic acid molecules encoding those polypeptides. Also provided herein are vectors and host cells comprising those nucleic acid sequences, chimeric polypeptide molecules comprising the polypeptides of the present invention fused to heterologous polypeptide sequences, antibodies which bind to the polypeptides of the present invention and to methods for producing the polypeptides of the present invention.

Secreted and Transmembrane Polypeptides and Nucleic Acids Encoding the Same

The present invention relates to the field glycosylation engineering of proteins. More particular, the present invention is directed to the glycosylation engineering of proteins to provide proteins with improved therapeutic properties, e.g., antibodies, antibody fragments, or a fusion protein that includes a region equivalent to the Fc region of an immunoglobulin, with enhanced Fc-mediated cellular cytotoxicity.

Glycosylation engineering of antibodies for improving antibody-dependent cellular cytotoxicity

https://www.kidney-international.org/article/S0085-2538(15)46731-6/pdf

Long-term survival in renal transplant recipients with graft function.

Following a diabetogenic T cell from genesis through pathogenesis.

The experience of the Peter Bent Brigham Hospital with 217 renal allografts functioning for more than 5 years is reviewed. Patient and graft survival were similar after 5 years, with patient survival being 88 and 66% at 10 and 15 years, respectively, and graft survival 85 and 75% at the same time intervals. Actuarial graft survival at 15 years was higher than patient survival because death with a functioning graft was not considered to be graft failure. No differences in patients or graft survival were found between living related and cadaver donor allografts. There were 33 deaths (15.2%), occurring from 5 1/2 to 20 1/2 years post-transplantation. Chronic liver failure and sepsis were the most common causes of death. Thirty-two patients (14.7%) lost their grafts after 5 years, most commonly from chronic rejection. Another 33 patients (15.2%) had evidence of graft dysfunction secondary to chronic rejection, recurrent glomerulonephritis, ureteral obstruction, or renal artery stenosis. Chronic rejection was generally not responsive to alterations in immunosuppressive medication. Complications of varying severity were common affecting 204 (94%) of the patients. The most frequent were hypertension, cataracts, avascular necrosis, malignancy, urinary tract infection, and pneumonia. These data demonstrate that transplant-related mortality and morbidity continue to occur in recipients of long-term renal allografts. These patients require careful and continuing care in medical centers experienced in transplantation.

Late mortality and morbidity in recipients of long-term renal allografts.

Liver disease in recipients of long-functioning renalallografts

Since the ftrst comprehensive description of skin graft rejection (Medawar 1944), much effort has been expended to clarify and define the remarkably complex and intricate host immunological responses called into play against foreign tissues. Knowledge has increased progressively in the past several years, especially with characterization of lymphocyte subsets and their influence upon one another. However, much of these data has been derived through experiments performed in vitro; the actual immunological events happening within a host stimulated by a histo-incompatibte graft, or those events occurring within the rejecting graft itself, remain poorly understood. In addition, although earlier dogma concerning the host responses were garnered primarily from skin allograft models, it has become increasingly clear that reactions to skin grafts are different from those toward organ grafts. During recent years, we have examined various cellular and humoral mechanisms occurring during acute rejection in unmodified animals as well as in tissues surviving for prolonged periods in immunologically manipulated hosts. The purpose of this review is to summarize some of the work done in our laboratories on particular aspects of host responses to vascularized organ allografts, primarily the heterotopic cardiac allograft transplanted between inbred strains of rats, but also failed human kidney grafts. Not only is the nature and function of cells accumulating within these allografts stressed, but the systemic nature of the immunologica! responses against a graft described by defining events occurring concomitantly in recipient iymphoid tissues. Because

Mechanisms of rejection and prolongation of vascularized organ allografts.

Mice sensitized with alloantigens and treated with cyclosporin A (CsA) were incapable of generating antigen-specific cytolytic lymphocytes (CL). Lymphocytes from these CsA-treated animals could not be reactivated upon exposure to the same alloantigens in mixed lymphocyte culture (MLC), whereas their response to a third-party antigen remained intact, suggesting a long-lasting and specific effect of CsA. After being irradiated, these lymphocytes from CsA-treated animals were added to normal MLC and were shown to prevent normal lymphocytes from becoming cytolytic in a dose-dependent and antigen-nonspecific fashion. These suppressor cells were not detected in mice receiving CsA only, indicating that CsA did not induce but rather permitted the expression of suppressor cells possibly generated by allosensitization. The suppressor cells appeared to be T lymphocytes, because treatment with anti-Thy-1.2 antibody and C abrogated their suppressive activity. The present results suggest that activation and/or sparing of suppressor cells by CsA may account for the long-lasting unresponsiveness seen in CsA-treated animals.

Evidence for the presence of suppressor T lymphocytes in animals treated with cyclosporin A.

Disclosed is a method of localized immunosuppression which may be used for preventing graft rejection or for preventing tissue destruction due to autoimmune disease. Also disclosed is a protein suppressor factor that is secreted by cloned anergic T-cells, blocks interleukin 2 (IL-2) stimulated T-cell proliferation, has an apparent molecular weight of between 10 and 30 kilodaltons, can be inactivated by heating to 65° C. for 15 minutes, blocks interleukin 4 (IL-4) stimulated T-cell proliferation in vitro, is non-cytotoxic to T-cells, and does not inhibit the production of IL-2 by T-cells in vitro.

Terry B. Strom

Papers

Late mortality and morbidity in recipients of long-term renal allografts.

Liver disease in recipients of long-functioning renalallografts

Mechanisms of rejection and prolongation of vascularized organ allografts.

Evidence for the presence of suppressor T lymphocytes in animals treated with cyclosporin A.

Methods and compounds for prevention of graft rejection