Karen Nelson Laboratory

The Nelson laboratory studies the response of CD8 T lymphocytes to donor human leukocyte antigens (HLA) after transplantation, identification of the elements generated from clinical laboratory assays that make up the profile of a recipient of a successful solid-organ transplant, and immunomodulation following transfusion.

CD8 T lymphocytes and Transplantation
T lymphocytes of the lineage identified by the CD8 marker have receptors for class I MHC plus peptide. Their role in response to transplanted organs has historically been thought to be limited to mediators of acute rejection via direct cytotoxicity or interactions between receptors and proteins of the tumor necrosis factor (TNF) family. However, the larger group of CD8 T cells includes cells with the capacity to down-regulate the activity of other T cells either via direct contact, via secreted cytokines, or via action on antigen-presenting cells. These CD8 cells with regulatory capacity can be differentiated by the expression of the cytokine IL-10 as opposed to the expression of granzymes or other cytotoxic mediators. Dr. Nelson’s lab has been analyzing peripheral blood samples taken prior to and following transplantation for CD8 cells that proliferate in response to re-exposure to donor HLA antigens ex vivo. The cytoplasm of proliferating cells has been stained to identify expression of IL-10, IFN-gamma or granzymes to identify cells with regulatory or cytotoxic phenotypes. The lab’s hypothesis is that increases in cells with regulatory capacity after re-stimulation with donor tissue is a marker for a reduced risk of rejection.

Transplant Profiles
The rapid translation of research techniques into clinically applicable assays and methods is enabling the lab to individualize the management of patients after transplantation. The assays employed allow snapshots of the immune response to HLA and tissue–specific antigens of the transplant donor. Data from these snapshots are then assembled to make up a profile that can be correlated with real-time data on the status of the transplanted organ. Elements associated with stable function or pending rejection will in the future be used to guide selection of immunosuppressive strategies or monitor tolerance-induction protocols.

Identification of Antibodies to HLA in Solid Organ Transplantation
Recent advances in microparticle-based flow cytometry assays now allow for the detection and identification of antibodies against single HLA molecules. Dr. Nelson’s lab is participating in research to identify the clinical correlates of different titers of antibodies detected by these assays: immune memory or active response. Data from these assays may form the basis for allocation of organs from deceased donors. The lab is partnering with other HLA laboratories in piloting this change. These new tools are not completely understood – there is ongoing research to identify artifacts and holes created by the nature of the recombinant molecules.

Data from these assays are also among the elements of the profile of an organ recipient. It was previously assumed that any donor-specific antibody generated after transplant was a predictor of rejection. Another focus of the lab’s studies is the hypothesis that antibodies with low titer or directed against antigens with a lower copy number may give survival signals and not mediate rejection.

Identification of Cytokine Profiles of Lymphocytes Responding to Donor Tissue
Lymphocytes responding to transplanted organs include the CD8 T lymphocytes described above, as well as CD4 T lymphocytes and natural killer t-cells (NKT) cells. Changes in these populations after transplant are elements of the organ recipient profile. NKT studies are performed in Dr. Warner’s laboratory.

Impact of Transfusion Component Processing
Filtration of transfusion components has been advanced as a solution to post-transfusion immunomodulation. However, most of the studies on allosensitization have used immunosuppressed subjects and studies on immunosuppression have used clinical outcomes as the endpoints. Dr. Nelson’s lab’s studies are targeting cardiac surgery patients (non-immunosuppressed) in collaboration with Dr. Gabriel Aldea at the University of Washington. The lab is measuring antibodies to HLA with the microparticle assays described above. These assays have the advantage of identifying changes in antibodies in patients previously allosensitized. To study immunosuppression, Dr. Paul Warner is analyzing the effects of transfusion on NKT cells and Yvette Latchman is studying CD4 T regulatory cells.

Transfusion-Associated Acute Lung Injury (TRALI)
In collaboration with Dr. Theresa Nester, the Blood Center’s Associate Medical Director, and Dr. Terry Gernsheimer, the Blood Center’s Director of Medical Education, Dr. Nelson’s lab is measuring antibodies to HLA antigens in transfusion components as well as in recipients with clinical symptoms meeting the criteria for TRALI.

1. Verghese PS, Smith JM, McDonald RA, Schwartz SM, Nelson KA,Warner PR. Impaired graft survival in pediatric renal transplantrecipients with donor-specific antibodies detected by solid-phase assays. PediatrTransplantation 2010: 14:730–734. PMID20598090
2. Kieran N, Wang X, Perkins J, Davis C, Kendrick E, Bakthavatsalam R, Dunbar N, Warner P, Nelson K, Smith KD, Nicosia RF, Alpers CE, Leca N, Kowalewska J. Combination of peritubularC4d and transplant glomerulopathy predicts late renal allograft failure. .J Am Soc Nephrol 10) :2260-8., 2009. PMID19729438
3. Slichter, S.J., Fish, D., Abrams, V.K., Gaur, L., Nelson, K., and Bolgiano, D. Evaluation of different methods of leukoreduction of donor platelets to prevent alloimmune platelet refractoriness and induce tolerance in a canine transfusion model. Blood, 105(2):847-854, 2005. PMID15231575
4. Reinsmoen, N., Zeevi, A., and Nelson, K.A. Anti-HLA antibody analysis and crossmatching in heart and lung transplantation. Transplant Immunol, 13: 63-71, 2004. PMID15203130
5. Gaur, L.K., Nitta, Y., Kennedy, E., Lernmark, A., Nelson, K.A., Allen, M., and Nepom, G.T. Induction of islet allotolerance in nonhuman primates. Ann N Y Acad Sci 958:199-203, 2002. PMID12021106
6. He, J.Q., Gaur, L.K., Stempien-Otero, A., Nelson, K., Levy, W.C., O’Brien, K.D., Bolgiano, D.C., and Reiner, A.P. Genetic variants of the hemostatic system and development of transplant coronary artery disease. J Heart Lung Transplant 21:629-36, 2002. PMID12057695
7. Nitta, Y., Nelson, K., Gaur, L.K., Andrews, R.G., and Allen, M.D. CFSE dye dilution mixed leukocyte reactions monitor alloreactivity following cardiac transplantation in non-human primates. Transplantation Proceedings, 33:226-9, 2001. PMID11266844
8. Reiner, A.P., Teramura, G., Nelson, K., and Slichter, S.J. A platelet monoclonal antibody-inhibition assay for detection of glycoprotein IIb/IIIa-related platelet alloantibodies. J Immunol. Meth.,184:153-162, 1995. PMID7658019
9. Nelson, KA., George, E., Swenson, C., Forstrom, J.W. and Hellstrom, K.E. Immunotherapy of murine sarcomas with auto-antiidiotypic monoclonal antibodies which bind to tumor-specific T cells. J. Immunol. 139:2110-2118, 1987. PMID2957449
10. Forstrom, J.W., Nelson, KA., Nepom, G.T., Hellstrom I., and Hellstrom, K.E. “Immunization to a syngeneic sarcoma by a monoclonal auto-anti-idiotypic antibody.” Nature 303:627-629, 1983. PMID6190090
11. Nelson, K., and Sjogren, H.O. “Effects of presensitization in vivo on cell-mediated responses to embryonic antigens in vitro.” Int. J. Cancer 10:108-114, 1978. PMID624596
12. Nelson, K., Pollack, S.B., and Hellstrom, K.E. “Specific anti-tumor responses by cultured immune spleen cells. I. “In vitro culture method and initial characterization of factors which block immune cell-mediated cytotoxicity in vitro.” Int. J. Cancer 15:806-814, 1975. PMID1140873

Mary S. Gallagher BS ,MT (ASCP), CHS (ABHI)
Research Associate
Email: [email protected]
Phone: (206) 292-3698
FAX: (206) 292-6582