The creation of long-lived HPA-1a specific T-cell clones

Abstract

Neonatal alloimmune thrombocytopenia (NAIT) is a disease where the fetus or newborn experience pathological low levels of blood platelets. This is a potentially fatal condition. Antibodies produced by the mother, crossing the placental barrier, targeting the β3-integrins expressed on fetal platelets, drive the disease. The antibodies are initially produced by B-cells, which are dependent on T-cells in order to mount a full response. Therefore, T-cells specific for the same antigen derived from the β3-integrin are believed to be vital in the pathogenesis of the disease. This makes these T-cells an obvious focus of research.

T-cells die in prolonged culture, and in this study three different strategies were used in an effort to obtain specific T-cell clones capable of sustained culture. This would be of great help in future research in our group. Human or murine T-cells with a known specificity were fused with the appropriate immortal partner cell line in order to create hybridoma cells. Further, human T-cells were transfected in order to express ectopic level of telomerase, in order to escape from cell senescence experienced upon multiple cell divisions, due to the shortening of their telomeres.

None of the methods described worked in full. The potential T-cells may serve as useful tools in the ongoing NAIT research in our lab. Either as standards used in the development of reagents capable of identifying specific T-cells isolated from patient sample, or in further detailing the peptide specificity driving the disease.