Epidermal cell derived factor (EDF): partial purification and characterization.
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Biomedical subjects
Publications and source records attributed to M Eisinger.
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Human epidermal cells grew and differentiated in vitro, provided that the pH of the culture medium was at 5.6-5.8, the seeding density was optimal (approximately 2.5 x 10(5) cells per cm2), and the incubation temperature was maintained at 35-37 degrees C. Under these conditions, epidermal cells from many different skin locations grew to confluency within 15-20 days and formed multi-layered sheets whose differentiated structure resembled that of the full depth of skin epidermis. Cell proliferation and differentiation did not require a feeder layer, a collagen substrate, a high concentration of fetal bovine serum, or added hormones. The sheets of differentiated epidermal cells could be dissociated from the plastic surfaces of the tissue culture flasks. The use of such cultured cells for wound dressing is proposed.
This report offers a description of a quantitative micro-complement fixation method (Cikes, 1975) for detecting human wart virus antigens and their specific antibodies, and proof of the specificity of the reactions being detected. The increased sensitivity demonstrated by chromium-release measurement is compared to the results of visual interpretation of complement fixation.
Virions isolated from warts of 2 siblings with epidermodysplasia verruciformis (EV), a rare disease characterized by the lifelong growth of warty skin tumors containing papova-like virions, were compared to isolates of human papillomavirus (HPV) from 3 pools of plantar and common hand warts. The length of relaxed, circular (form II) molecules of EV virion DNA approximated the length of HPV DNA but was 3.3% longer. Antisera prepared in rabbits against the 3 HPV pools coated and aggregated HPV in immune electron microscopy (IEM) tests but did not react with EV virions. These antisera reacted at high titers in complement fixation (CF) tests with HPV and reacted only weakly in CF tests with EV virions. Rabbit antisera to EV virions coated and aggregated EV virions but reacted only weakly or not at all with HPV virions in IEM tests. These sera reacted in CF with EV virions only. The data indicated that virions from the EV patients represent an immunologically distinct papillomavirus.
Lymphocyte transformation (LT) and leucocyte migration inhibition in agarose were used to demonstrate cell-mediated immune response to purified human wart virus and wart tissue extract in various subjects with warts and those without past history of warts. Most individuals bearing warts for less than 1 year duration showed positive cell-mediated responses to both the virus and tissue extract whereas very few of those who had warts for longer duration responded to either antigenic preparation. The difference was statistically significant. Subjects who had warts in the past also showed positive responses but these tended to decrease in degree with time. Surprisingly a group of subjects who never had warts before also responded to stimulation with the virus, but not to the extract. The positive response to stimulation with wart tissue extract reflects the presence of wart associated antigens other than the virus. Cell-mediated immunity against the wart virus and wart-associated antigens is probably important in preventing the persistence or even establishment of disease but this protective immunity is short-lived. The lack of quantitative correlation between LT and leucocyte migration inhibition demonstrable in this study suggests that these two are separate events in in vitro lymphocyte stimulation with antigens.
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