A SIMPLE METHOD FOR THE PROPAGATION OF CERVICAL LYMPHOCYTES

Local immune function is most likely a key influence in the establishm ent of human papillomavirus infections and its subsequent disease, Unf ortunately, little information is known about local cervical immunity, and even less is known about human papillomavirus immunoreactivity. I n addition, studies of local immunoreactivity have been hampered by th e technical difficulty in obtaining cervical lymphocytes. The objectiv e of the present study was to develop a simple method for the propagat ion of cervical lymphocytes from biopsy-size specimens, Cervical tissu e was obtained from women undergoing a hysterectomy. Cervical samples measuring approximately 3 by 5 by 2 mm were minced and divided into tw o portions, One portion was digested by standard digestion methods and density gradient lymphocyte separation, The sample was then immuno-ch aracterized for CD4 and CD8 cells by how cytometry, The other portion was minced into 1-mm(3) sections, and each section was placed into a s eparate well with tissue culture medium and interleukin 2. Lymphocyte counts and immunophenotypic analysis were performed after 18 to 20 day s in culture, After 18 to 20 days in culture, the analysis demonstrate d that this method of direct lymphocyte culture from a biopsy specimen yielded approximately 1 x 10(6) to 5 x 10(6) lymphocytes. Immunopheno typic studies of the digested sample at day 0 revealed CD4-to-CD8 rati os of between 0.7:1 and 3.5:1, and at days 18 to 20 they revealed rati os of between 2.3:1 and 98:1, In summary, we developed a simple techni que for propagating cervical lymphocytes from small tissue samples for the study of the local immune response, Studies are under way to opti mize lymphocyte growth and to preserve CD8 populations.