Sodium caseinate induces mouse granulopoiesis.

OBJECTIVE AND DESIGN: Sodium caseinate (CasNa) induces differentiation and M-CSF production in mouse band granulocytes in vitro; however, it is not yet known if this molecule can also induce the proliferation and activation of the granulocyte lineage in vivo. In this work we evaluated the induction in vivo of granulopoiesis and the activation of granulocytes in mice treated with CasNa. MATERIAL OR SUBJECTS: BALB/c male mice 8-12 weeks old were used. TREATMENT: The animals were inoculated intraperitoneally with 1 ml of CasNa (10% in PBS p/v) four times (every 48 h). METHODS: Granulocyte proliferation was evaluated by flow cytometry; activation was evaluated by phagocytic indices. The cytokine was measured using an ELISA assay. RESULTS: We show that CasNa increased bone marrow granulopoiesis percentage (38.35 ± 10.88 vs. 64.94 ± 34.14 BrdU+/Gr-1+ cells) and the granulocytes generated presented increased phagocytic indices (0.3 ± 0.1 vs. 0.6 ± 0.11, p < 0.05). We also show that G-CSF (974 ± 411 vs. 3189 ± 350 pg/ml, p < 0.05) and GM-CSF increased in serum, but only G-CSF in bone marrow plasma. CONCLUSIONS: CasNa induces granulopoiesis with functional granulocytes, suggesting that this molecule could be an innate immune system activator.

GST-pi expression in BCR-ABL+ and BCR-ABL- cells from CML patients.

Our laboratory has been involved in the study of glutathione-sulfhydryl-transferase-pi (GST-pi) for several years. We have recently observed that during haematopoiesis in BMSC liquid cultures from CML patients who were candidates for transplant GST-pi was expressed in presumably malignant cells during different stages of cellular maturation. To confirm this finding, in the present work we are detecting GST-pi expression by immunofluorescence in BCR-ABL+ and BCR-ABL- cells done by FISH of PB from 30 CML patients during different clinical status: treatment (T), hematological relapse (R), blastic crisis (BC) or post-allotrasplant (PT). As well as in PB from 30 Blood-Bank donors. The results were %BCR-ABL+ GST-pi+ cells: T = 1-67, R = 33-69, BC = 90-100 and PT = 1-2; %BCR-ABL- GST-pi+ cells: T = 2-31, R = 5-18, BC = 0-10 and PT = 2-5; %BCR-ABL- GST-pi- cells: T = 2-97, R = 13-62, BC = 0 and PT = 93-96; %BCR-ABL+ GST-pi- cells: T = 0, R = 0, BC = 0 and PT = 0. GST-pi was not expressed in donor cells. The results obtained confirm our previous observations and suggest that GST-pi expression might be used for the evaluation of the minimal residual disease in CML patients.

Follow-up of GST-P during hepatocarcinogenesis with DEN-2AAF in F344 rats.

Although it is known that doublets of hepatocytes GST-P+ are produced during the first week after initiation with DEN, the activity levels of the enzyme are not known in the initial stages of the process, neither is its behavior through an initiation-promotion scheme. We consider the latter issues important in order to obtain information of the initiation step and its relation with GST-P.DEN was applied as initiator in a single dose on day 0 to F344 rats (200 mg/kg) and as promoter 2-AAF in 20 mg/kg doses on days 7, 8 and 9 of the scheme. Partial hepatectomy was performed on day 10, and daily during the 28 days in which the experiment lasted. The GST-P activity was determined in postmicrosomal supernatants of respective livers (by immunoadsorption) as well as their histological section (by immunohistochemistry). In both cases antibody anti-GST-pi produced in our laboratory was employed. The results obtained show GST-P appearance on day 5 of the scheme in rats treated with carcinogens. The activity of the enzyme increased slowly reaching its maximum on day 18, together with the appearance of GST-P+ preneoplastic nodules. Our results suggest that GST-P could display an additional function to those previously known, in cellular differentiation this could explain the very frequent expression of this enzyme in preneoplastic as well as in neoplastic cells.