Fractionation of functional lymphocytes sensitized to basic encephalitogen on derivatized collagen and gelatin gels.
The lymph node cells of basic encephalitogen (BE)-sensitized guinea pigs were fractionated on derivatized collagen and gelatin gels. The population of cells specifically reactive to this antigen can be isolated from derivatized gelatin gels and retain their viability and functionality as assayed in vitro. The specific binding of BE-sensitized cells to BE-derivatized gels comprised between 1 and 2% of the cells applied per plate. The ratio of sensitized cells bound to non-sensitized cells bound ranged between 4 and 6. The viability and functionality of adherent cells detached from collagen gels after enzymatic degradation were impaired. In contrast, the responses obtained with the adherent cell population released from the gelatin gels, by melting at 37 degrees C, were equal or greater than those of the original unfractionated lymph node cell cultures. Furthermore, it was possible to obtain a nonadherent cell population which was virtually completely depleted of the capacity to respond to the sensitizing antigen.
D000818:Animals; D001665:Binding Sites; D002250:Carbon Radioisotopes; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D002478:Cells, Cultured; D003094:Collagen; D003851:Deoxyribonucleases; D005780:Gelatin; D005782:Gels; D006168:Guinea Pigs; D007114:Immunization; D008198:Lymph Nodes / Q000166:cytology; D008213:Lymphocyte Activation; D008212:Lymphocyte Depletion; D008214:Lymphocytes / Q000276:immunology*; D003012:Microbial Collagenase; D009113:Muramidase; D004676:Myelin Basic Protein; D013936:Thymidine / Q000378:metabolism
D002250:Carbon Radioisotopes; D005782:Gels; D004676:Myelin Basic Protein; D005780:Gelatin; D003094:Collagen; D003851:Deoxyribonucleases; D009113:Muramidase; D003012:Microbial Collagenase; D013936:Thymidine