cytotoxic activity of various herbals against different tumor cells: an in vitro study

Medicinal plants have been investigated for possible anti-cancer effects. The aim of the present study was to examine the cytotoxic activity of several medicinal native plants on different tumor cell lines. Plants including Salvia santolinifolia, Salvia eremophil, Salvia macrosiphon, Salvia reuterana, Teucrium persicum, Anvillea gracini and Francoeuria undulate were collected from different sites of Fars Province in southern Iran. The methanolic extracts of the plants were prepared and their effects at concentrations of 5-200 micro g/ml on various tumor cell lines were examined using a colorimetric assay. Among the extracts of Salvia species, the strongest inhibitory effect was observed for S. reuterana extract. This extract showed a strong cytotoxic effect on the Raji lymphoma cell line. More than 50% of Raji cells growth was inhibited by 21 micro g/ml of this extract. S. macrosiphon extract also showed a strong inhibitory effect on this tumor cell line [IC50=77 +/- 1 micro g/ml]. The greatest inhibitory effect of T. persicum extract was on Hela tumor cell line. This extract at concentration of 69 +/- 2 micro g/ml causes 50% inhibition of Hela cell growth. Corresponding data for A. gracini extract was obtained at concentrations of 83.5 +/- 2 and 86 +/- 3 micro g/ml on Jurkat and Hela cells, respectively. F. undulata reduced the proliferation of all the tumor cell lines used in this study but this inhibition did not reach 50%. All the extracts, more and less, showed inhibitory effects on the tumor cell lines. The most cytotoxic activity was observed in S. reuterana with an IC50 value less than 25 micro g/ml towards Raji cell line

Determination of anti-ds-DNA antibody levels in patients with systemic lupus erythematosus by Elisa, RIA and crithidia methods: Changes in antibodies prior to disease activity

Antibodies [Abs] to double-stranded [ds] DNA have been considered most useful in diagnosis and monitoring disease activity in patients with systemic lupus erythematosus [SLE]. To investigate the value of anti-ds-DNA Ab level changes, detected by three different methods, as predictor of disease activity in SLE patients. Serial sampling from 87 patients with SLE was performed. Samples were assessed for anti-ds-DNA Abs by Crithidia luciliae test [CLT], ELISA and Farr assay. In 17 out of 40 cases with completed sampling, disease activity was occurred during the study. Significant increases in anti-ds-DNA Ab level was detected by ELISA [whole and IgG] and Farr assay prior to disease activity in all clinically active patients. CLT was tested positive in only one case before disease activity. None of the 23 cases with clinically inactive disease were tested positive with CLT, whereas in 3 and 15 of 23 cases positive results were observed by ELISA and Farr assay, respectively. Statistical analysis showed the predictive value of ELISA for disease activity. There was a high correlation between the results of Farr assay and ELISA for all SLE patients in detecting anti-ds-DNA Abs [r=0.63, p<0.0001]. ELISA and Farr assay were also correlated well with disease activity [p<0.05]. Our findings confirm the predictive value of increases in anti-ds-DNA Abs measured by ELISA method with respect to activity of SLE

Immunophenotypic analysis in Iranian patients with acute myelogenous leukemia: correlation with leukemia subgroups

Immunophenotyping has led to important insights into leukocyte differentiation and cellular origin of leukemias. This study was undertaken to analyze the expression of certain leukocyte surface markers in acute myelogenous leukemia and to find its possible correlation with a morphological classification. Methods:myelogenous The immunophenotypic characteristics of 70 patients with diagnosis of acute leukemia [AML] were investigated using a panel of monoclonal antibodies [mAbs] against CD13, CD14, CD15, CD33, CD34, CD38, CD71, CD11b and HLA-DR molecules. The frequency of expression of each surface marker in relation to morphological classification by French-American-British [FAB] criteria was analyzed. Among the mAbs used in this study HLA-DR, CD38 and CD15 were positive in the majority of cases. CD15, CD33, CD34 and CD13 were the most common antigens expressed in M1 patients. In M2 subtype, the highest expression belonged to CD13 molecule. The blast cells of all 9 patients with M3 AML expressed CD15 antigen. In these patients CD38 had the least expression in comparison with other subtypes. Blast cells of the largest group [M4] showed high reactivity with CD11b and CD15 mAbs. In M5 and M6 patients HLA-DR, CD38, CD15, CD11b [for M5 group] and CD71 [for M6 group] were found in all cases. CD14, which had an expression of 75% in M5 cases, was practically absent in other subtypes. Some immunophenotypes correlated with FAB type, including increased frequency of CD71 expression and lack of expression of CD11b in M6 group [p<.05], lack of expression of CD71 and decreased expression of CD38 in M3 patients [p<.05], and increased frequency of CD13 and CD14 expression in M2 and M5 groups respectively [p<.008]

Expression of HLA-class I and II, ICAM-1 and CD44 antigens in transitional csll carcinoma of bladder

The expression of monomorphic determinants of the histocompatibility leucocyte antigens [HLA] class I and class II, ICAM-1 and CD44 were studied using immunochemical staining of bladder tissue specimens from 30 cases of transitional cell carcinoma [TCC] of bladder. The results indicated that HLA-class I antigens were relatively well preserved in low grade cases but high grade tumors, either lacked or had markedly reduced HLA-class I antigens. Statistical analysis showed a significant difference between tumor grade in the expression of these molecules [p<0.05]. Aberrant expression of HLA-DR was detected in 30% of cases and moderate to strong expression of ICAM-1 and CD44 molecules were observed in 43% and 50% of cases, respectively. No significant difference in ICAM-1 and CD44 expression was found in relation to differentiation of tumor cells

Identification of two different intracellular antigens in human leukocyte by monoclonal antibodies

Two monoclonal antibodies [mAbs] designated as SU11 and SUI2 were produced by fusion of a mouse myeloma cell line with spleen cells from mice immunized with human tonsillar lymphocytes. SUII and SUI2 monoclonal antibodies recognized two epitopes with molecular weights of 39 and 43 KDa derived from solubilized membrane of Nalm -16 and Bjab cell lines, respectively. Flow cytometric analysis revealed that these mAbs were reactive to around 30% of peripheral blood T cells and 25% of B cells and granulocytes, Nuclear and cytoplasmic staining was observed in 90% of bone marrow myeloid and lymphoid cells. These characteristic features of SUI1 and SUI2 mAbs suggest that the epitopes recognized by these mAbs are scattered throughout the cell membrane cytoplasm and nucleus

CD38 molecule: a multilineage glycoprotein and its unique expression on plasma cells

A hybridoma clone designated 6G5 has been selected by fusion of mouse myeloma cell line Ag. 8653 with spleen cells from mice immunized with human peripheral blood mononuclear cells [PBMC]. The antibody produced by this clone was found to be strongly reactive with four human B-cell lines in the conventional immunological assays. Despite the fact that expression of most B cell-associated markers are lost upon differentiation of B-cells to plasma cells, the expression of the 6G5 reactive molecule remains unchanged. The lack of reactivities of this MAb for mature T-cells, and monocytic cell lines indicates that this MAb recognizes a B cell associated marker. Western blot analysis indicated that the 6G5 MAb detected a single band with molecular weight of41KDa from cell lysates of two human B-cell lines, DAUDI and NALM6. Comparison of data obtained for 6G5 MAb with those of the MAb known as OKT10 indicated that both MAbs may have reacted with the same molecule

production of murine monoclonal antibodies [MAb] directed against human T-lymphocyte subsets

The production of murine monoclonal antibody [MAb] has not yet been reported in Iran. The present work describes for the first time the generation of several murine hybridoma clones secreting MAbs directed against human leukocyte surface antigens. The secreted antibodies by hybridoma clones have been screened on different lymphoid and non-lymphoid tissues. Results indicated that of seven hybridomas, clones 3F11, 3C3 and 1F2 showed a strong reactivity for T-cells purified from thymus and tonsil tissues. Moreover, clones 1 D4 and 6G5 which partially stained thymus tissues were found to be negative on purified B-cells and monocytic cell line U937. The third group, 4E5 and 4F4 hybridoma clones was expressed weakly on purified T, B-cells and U937 cell line. Further work is needed to determine the epitopes recognized by these MAbs and a comparison of the data with those presented at the previous leukocyte antigens workshop