Assuntos
Anticorpos/isolamento & purificação , Células Produtoras de Anticorpos/imunologia , Animais , Especificidade de Anticorpos , Ensaio de Unidades Formadoras de Colônias , Eritrócitos/imunologia , Retroalimentação , Feminino , Técnicas de Imunoadsorção , Terapia de Imunossupressão , Técnicas In Vitro , Camundongos , Camundongos Endogâmicos CBA , Staphylococcus aureus/imunologiaAssuntos
Medula Óssea/imunologia , Tecido Linfoide/imunologia , Animais , Formação de Anticorpos , Anuros , Cobaias , Técnica de Placa Hemolítica , Contagem de Leucócitos , Leucócitos/imunologia , Linfócitos/imunologia , Mitose , Rana catesbeiana , Rana pipiens , Baço/imunologia , Fatores de Tempo , XenopusRESUMO
A miniaturized diffusion culture apparatus holding 24 individual cultures and utilizing 1/10 the cells and volume of standard Marbrook-type cultures is described. This system was evaluated by studying the anti-sheep erythrocyte response of spleen cells from unprimed or primed and boosted mice. Compared to parallel Mishell--Dutton plate cultures memory responses in mini-diffusion cultures rise more slowly, peak 1--2 days later and thereafter decline less rapidly. Moreover, the diffusion cultures give a pronounced IgG response peaking at 6 days, while the abortive IgG response evident in plate cultures at 4 days rapidly disappears. While viability in the two systems is equivalent, cell recovery on days 7 and 8 is markedly higher in the diffusion cultures. Of 4 cell concentrations tested, 2 million cells are found to produce the optimum anti-sheep erythrocyte plaque-forming cell response. A 0.2 micron nucleopore membrane is preferable to a dialysis membrane and yields up to a 2-fold higher response. Replacement of the reservoir medium on day 4 impairs rather than improves the response. The culture system was also found to support an excellent primary response. This response is dependent on the presence of 2-mercaptoethanol and does not require rocking.
Assuntos
Linfócitos/imunologia , Animais , Sobrevivência Celular , Células Cultivadas , Relação Dose-Resposta Imunológica , Eritrócitos/imunologia , Feminino , Técnicas Imunológicas , Membranas , Mercaptoetanol/farmacologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos CBA , OvinosRESUMO
An ear assay was used to detect delayed responses in mice. Animals were sensitized and subsequently injected with 3H thyminedeoxyryribose to label antigen sensitive cells. Each animal was given a control and a test ear challenge and the respective responses were assessed by scintillation counting of 3H in cells which had accumulated at the reaction site. Delayed responses were consistently observed in animals sensitized to dinitrochlorobenzene (DNCB) but not in animals sensitized to sheep red blood cells. Reactivity to DNCB was minimal in tolerant animals and animals sensitized to the control agent. Both peritoneal exudate and spleen cells of antigen-sensitized animals were found to contain more label than cells from non-sensitized animals.
