Walker 256 tumor MHC class I expression during the shift from A variant to the immunogenic AR variant.

Novel tumor cell variants can be obtained by serially passaging tumor cells in different media and/or environments. Serial intraperitoneal (ip) passages of the Walker 256 tumor A variant was followed for studying the generation of its regressive AR variant. MHC class I molecule expression was assessed since variations in this molecule would explain changes in tumor cell immunogenicity and therefore, the shift from progressive A variant to the regressive AR variant. Within 25 ip passages all serial repetitions shifted from A to AR variant, which was characterized by a significant increase in red blood cell (RBC) osmotic fragility with marked spleen hypertrophy in the host. In one serial repetition AR tumor cells were rejected (ip passage number 36) and immunity against the AR and A variants was conferred. Flow cytometry analysis showed a significant increase in the number MHC class I positive cells in AR variant (n = 15, 14.21 +/- 1.32) compared with A variant (n = 10, 9.10 +/- 1.22). These data provide evidence that the generation of the AR variant could result from factors present in the ip environment leading to an increase in the number of Walker 256 MHC class I positive tumor cells, probably due to immune selection of MHC class I negative tumor cells.

Tumor growth characteristics of the Walker 256 AR tumor, a regressive variant of the rat Walker 256 A tumor

The present study aimed at characterizing the subcutaneous development of the Walker 256 (W256) AR tumor, a regressive variant of the rat W256 A tumor. Wistar rats were injected subcutaneously with 4x10(6) W256 A or W256 AR tumor cells. The development of tumors was evaluated daily by percutaneous measurements. None of the W256 A tumors (n=20) regressed, but 62 percent of the W256 AR tumor-bearing rats (n=21) underwent complete tumor regression within 35 days. Continuous growth of AR tumors was characterized by an increase of the tumor growth rate from day 12, which reached values above 1.0 g/day, and were significantly higher (p<0.05) than those of the regressive AR tumors. Immunosuppression by irradiation before subcutaneous injection of AR cells completely abrogated tumor regression and was associated with severe metastatic dissemination. Daily evaluation of the tumor growth rate enabled the discrimination, in advance, between continuously growing tumors and those that regressed later on.

O objetivo neste estudo foi caracterizar o desenvolvimento subcutâneo do tumor de Walker 256 (W256) AR, uma variante regressiva do tumor de W256 A de rato. Ratos Wistar foram injetados com 4x10(6) células tumorais de W256 A ou W256 AR. O desenvolvimento tumoral foi avaliado diariamente. Nenhum dos tumores W256 A (n=20) regrediu, mas 62 por cento dos ratos com tumor W256 AR apresentaram regressão completa dos tumores em até 35 dias. O crescimento contínuo dos tumores AR foi caracterizado pelo aumento da taxa de crescimento tumoral a partir do dia 12, alcançando valores maiores que 1,0g/dia, que foram significativamente superiores (p<0,05) aos valores de taxa de crescimento dos tumores regressivos AR. A imunossupressão por irradiação precedendo a injeção das células tumorais AR eliminou completamente a regressão tumoral e favoreceu disseminação metastática severa. Este estudo caracterizou o desenvolvimento do tumor de W256 AR em condições específicas, documentando a regressão espontânea deste tumor após a injeção subcutânea de altas doses de células tumorais em ratos Wistar. A avaliação diária da taxa de crescimento tumoral permite discriminar precocemente os tumores com crescimento continuo daqueles que são regressivos. A taxa de crescimento tumoral é um parâmetro útil para a avaliação dos animais experimentais, particularmente no período que precede a regressão dos tumores.