Proliferation Of Human Peripheral Blood Lymphocytes Research Articles

Performance evaluation of two antigen-extracted xenogeneic ostein and experimental study on repairing skull defects in rats

To evaluate the physical and chemical properties, immunogenicity, and osteogenesis of two antigen-extracted xenogeneic bone scaffolds-decalcified bone matrix (DBM) and calcined bone. By removing the inorganic and organic components of adult pig femus, xenogeneic DBM and calcined bone were prepared respectively. The density and pH value of the two materials were measured and calculated, the material morphology and pore diameter were observed by scanning electron microscope, and the surface contact angle was measured by automatic contact angle measuring instrument. The safety, osteogenic activity, and immunogenicity of the two materials were evaluated by cytotoxicity test, osteoblast proliferation test, DNA residue test, and human peripheral blood lymphocyte proliferation test. The two materials were implanted into the 5 mm full-thickness skull defect of 6-week-old male Sprague Dawley rats (the blank control group was not implanted with materials). The materials were taken at 4 and 8 weeks after operation, the repair effect of the materials on the rat skull was observed and evaluated by gross observation, Micro-CT scanning, and HE staining observation. Compared with calcined bone, DBM has lower density and poor hydrophilicity; the pH value of the two materials was 5.5-6.1, and the pore diameter was 160-800 μm. The two materials were non-cytotoxic and could promote the proliferation of osteoblasts. The absorbance ( A) values of osteoblast proliferation at 1, 4, and 7 days in the DBM group were significantly higher than those in the calcined bone group ( P<0.05). The DNA residues of the two materials were much lower than 50 ng/mg dry weight, and neither of them could stimulate the proliferation and differentiation of human peripheral blood lymphocytes. The results of animal experiments in vivo showed that the bone volume/total volume (BV/TV) in DBM group and calcined bone group were significantly higher than that in blank control group at 4 weeks after operation ( P<0.05), and that in calcined bone group was significantly higher than that in DBM group ( P<0.05); at 8 weeks after operation, there was no significant difference in BV/TV between groups ( P>0.05). HE staining showed that at 4 and 8 weeks after operation, the defect in the blank control group was filled with fibrous connective tissue, the defect was obvious, and no bone growth was found; the defect in DBM group and calcined bone group had been repaired to varying degrees, and a large number of new bone formation could be seen. The material degradability of DBM group was better than that of calcined bone group. The physical and chemical properties and degradability of the two kinds of xenogeneic bone scaffolds were slightly different, both of them have no immunogenicity and can promote the repair and reconstruction of skull defects in rats.

Hepatoprotective Activities of Colocasia esculenta (L.) Schottin Mice Model with Liver Injury Induced by Paracetamol

This paper uses the model of experimental mouse liver damage with paracetamol to evaluate the liver protective effects of the methanol extract (Colocasia esculenta (L.) Schott) (MCE). After 8 days, the hepatoprotective activities of MCE in mice (damaged by paracetamol) at doses of 500, 1,000 and 2,000 mg/kg/day proved effective as the AST enzyme content decreased by 6.98, 22.84 and 26.59%, respectively; and ALT decreased by 53.17, 56.46 and 57.93%, respectively. MCE at a dose of 1,000 mg/kg/day had a protective effect equivalent to that of silymarin (used in liver treatment) at a dose of 50 mg/kg/day and MCE at a dose of 2,000 mg/kg/day had better effect. Observation of the microscopic liver tissue cross section also revealed that the mice treated with MCE of C. esculenta at doses of 1,000 and 2,000 mg/kg/day showed significantly improvement in their liver tissues compared to the non-treated control group.&#x0D; Keywords:&#x0D; Colocasia esculenta, hepatoprotective, paracetamol.&#x0D; References&#x0D; [1] S. Shahani. Evaluation of hepatoprotective efficacy of APCL-A poly herbal formulation in vivo in rats, Indian Drugs. 36(1999) 628-631.[2] Y. Cui, X. Yang, X. Lu, J. Chen, Y. Zhao. Protective effects of polyphenols-enriched extract from Huangshan Maofeng green tea against CCl4-induced liver injury in mice, Chemico- Biological Interactions 220 (5) (2014) 75-83. https://doi.org/10.1016/j.cbi.2014.06.018 [3] K.C. Choi, W.T. Chung, J.K. Kwon, J.Y. Yu, Y.S. Jang, S.M. Park, S.Y. Lee, J.C. Lee. Inhibitory effects of quercetin on aflatoxin B1 induced hepatic damage in mice, Food Chemical Toxicology, 48 (10) (2010) 2747-2753 . https://doi.org/10.1016/j.fct.2010.07.001[4] E.S. Sabina, J. Samue, S.R. Ramya, S. Patel, N. Mandal, P. Preety, P.P. Mishra, M.K. Rasool. Hepatoprotective and antioxidant potential of Spirulina fusiformis on acetaminophen induced hepatotoxicity in mice, International Journal of Integrative Biology 6 (1) (2009) 1-5. http://ijib.classicrus.com/.../1501.pdf[5] Pham Hoang Ho. Vietnamese plants, episode III. Young Publishing House, (1999), pp. 353 (in Vietnamese).[6] C.O. Eleazu, M. Iroaganachi, K.C. Eleazu. Ameliorative potentials of cocoyam (Colocasia esculenta L.) and unripe plantain (Musa paradisiaca L.) on the relative tissue weights ofstreptozotocin-induced diabetic rats. Journal of Diabetes Research, 197 (2013), https://doi.org/10.1155/2013/160964.[7] R.C. Cambie, L.R. Ferguson. Potential functional foods in the traditional Maori diet. Mutation Research Letters 523–524 (2003) 109-117. https://doi.org/10.1016/s0027-5107(02)00344-5[8] C.L. Abraham, K. Yoshinori, T. Masakuni, I. Hironori, O. Hirosuke, T. Hajime. Flavonoid glycosides in the shoot system of Okinawa Taumu (Colocasia esculenta S). Food Chemistry 119 (2010) 630- 635. https://doi.org/10.1016/j.foodchem.2009.07.004[9] P.J. Bouic, D. Etsebeth, R.W. Liebenberg, C.F. Albrecht, K. Pegel, J.P.P. Van. Beta sitosterol and beta sitosterol glycoside stimulate human peripheral blood lymphocyte proliferation: Implications for their use as an immunomodulatory vitamin combination. International Journal of Immuno-pharmacology 18 (12) (1996) 693-700. https://doi.org/10.1016/S0192-0561(97)85551-8.[10] Vo Van Chi. Dictionary of Vietnamese medicinal plants. Medical Publishing House (1997), pp. 871-873 (in Vietnamese).[11] Do Trung Dam. Method of determining drug toxicity. Medical Publishing House, Hanoi (2014), pp. 11-137 (in Vietnamese). [12] J.T. Litchfield, F. Wilcoxon. A simplified method of evaluating dose-effect experiments, Journal of Pharmcology and Experimental Therapeutics, 96 (2) (1949) 99-113. PMID: 18152921 [13] Doan Thi Nhu, Do Trung Dam, Pham Duy Mai. Method research of the pharmacological effect of drugs from medicinal herbs, Science and Technics Publishing House, Hanoi (2006), pp. 142 (in Vietnamese).[14] A. Godwin. Histochemical uses of haematoxylin - A Review. JPCS 1 (2011) 24-34. [15] Ho Thi Thanh Huyen, Thai Nguyen Hung Thu, Nguyen Thai An. Some results of plan microscopic studies and chemical composition of Bombax malabaricum DC., Bombacaceae, Proceeding Pharma Indochina VII, 10/2011, The 7th Indochina Conference on Pharmaceutical science, Bangkok, Thai Lan (2011) 270-274.[16] A.N. Chinonyelum, A.P. Uwadiegwu, O.C. Nwachukwu, O. Emmanuel. Evaluation of hepatoprotective activity of Colocasia esculenta (L. Schott) leaves on thioacetamide-induced hepatotoxicity in rats. Pakistan Journal Pharmaceutical Sciences, 28 (6S) (2015) 2237-2241. [17] O.F. Dayo, B.C. Eluke, E.O. Ukaejiofo, O.L. Olayinka, C.I. Johnpaul, O.A.K .Tajudeen, I. Chinwe, L.S. Adetona. Hepatoproactive and haematopoietic modulatory efficacy of leaf extract of Colocasia esculenta in Albino wistar rats. International Journal of Tropical Medicine 12 (3-6) (2017) 35-41. https://doi.org/3923/ijtmed.2017.35.41.&#x0D; &#x0D; &#x0D; &#x0D; &#x0D;

Suppression of human peripheral blood lymphocyte proliferation by immortalized mesenchymal stem cells derived from bone marrow of Banna Minipig inbred-line

This study sought to investigate whether mesenchymal stem cells (MSC) derived from Banna Minipig Inbred-line (BMI-MSC) suppressed human peripheral blood lymphocyte (hPBLs) proliferation in a one-way mixed lymphocyte reaction system. BMI-MSC failed to stimulate proliferative responses by hPBLs, which were activated by allogenic endothelial cells, BMI-PBLs and non-specific mitogenic stimuli. Furthermore, BMI-MSC also suppressed proliferation of hPBLs stimulated by mismatched allogenic, as well as xenogenic PBLs, and the mitogenic stimulus ConA. The suppression occurred in dose-dependent fashion when the ratio of hPBLs to BMI-MSC varied from 1 to 5 fold; fewer, BMI-MSC (0.001 to 0.01 times) showed no obvious suppression. When BMI-MSC were added to hPBLs stimulated for 72 hours, the proliferative suppression was still evident. Addition of anti-FasL or anti-TGF-β1 antibody attenuated the proliferative suppression, while antibody against IL-10 had no effect on it. Further immunofluorescence analysis demonstrated that FasL and TGF-β1 constitutively expressed BMI-MSC. These findings suggest that BMI-MSC suppress hPBLs proliferation relying on FasL and TGF-β1 mediated pathways.

SUPPRESSION OF HUMAN PERIPHERAL BLOOD LYMPHOCYTE PROLIFERATION BY IMMORTALIZED MESENCHYMAL STEM CELLS FROM AN INBRED PORCINE BREEDING

P1100 Aims: Use mesenchymal stem cells (MSC) from Banna Minipig Inbred Line (BMI) (the 18th generation of subline JS) to investigate its suppression effects on human peripheral blood lymphocyte proliferation and explore the possible mechanism underlying. Methods: MSCs from BMI (BMI-MSC) were immortalized by introducing simian virus 40 large T antigen gene (SV40 gene). Immunogenic and immunosupressive properties of BMI-MSC were investigated through one-way mixed lymphocyte reactions (MLR) using MTS assay to measure the proliferative activity of hPBLs. Results: BMI-MSC failed to elicit a proliferative response when cultured with hPBLs (Fig.1) and the viability of stimulated hPBLs were suppressed in a dose dependent fashion after 1 to 5 times xenogenic BMI-MSCs were added to ConA, PHA, allogeneic or xenogenic PBLs stimulated hPBLs(Fig.2). Furthermore, addition of anti-FasL or anti-TGF-β1, but not IL-10, could attenuated the inhibition activity of BMI-MSC in a dose dependent fashion(Fig.3). The expression of FasL and TGF-β1 of BMI-MSC were confirmed by Immunofluorescence analysis under laser confocal microscope (LCM).Figure 1. BMI-MSC failed to elicit a proliferative response when cultured with xenogenic hPBLs.Figure 2. Dose dependence inhibitory effect of immortalized BMI-MSCs on hPBLs proliferation induced by 10μg/ml ConA or 0.5%PHA.Figure 3. Modulation of PBLs proliferation by antibodies reacting with MSC derived cytokines.Conclusions: Our data suggested that BMI-MSC have significant immunosupressive effect on xenogenic hPBLs proliferation which might partially due to FasL and TGF-β1 mediated apoptosis. Engineered BMI-MSC might be a protencial seed cell in tissue engineering and immunomodulator in hematopietic as well as solid organ transplantation.

Beta-sitosterol and beta-sitosterol glucoside stimulate human peripheral blood lymphocyte proliferation: Implications for their use as an immunomodulatory vitamin combination

The phytosterols, β-sitosterol (BSS), and its glucoside (BSSG) enhance the in vitro proliferative response of T-cells stimulated by sub-optimal concentrations of phytohaemagglutinin (PHA) several fold at extremely low concentrations (femtogram level). A 100:1 (mass:mass) ratio of BSS:BSSG (termed essential sterolin formulation, ESF) showed higher stimulation than the individual sterols at the same concentration. In vivo activity of ESF was also demonstrated when volunteers ingested ESF for 4 weeks. Proliferation of their T-cells, stimulated maximally with PHA, was significantly enhanced (20–920%) when compared to baseline values. In vitro, ESF (1 μg/ml) was able to significantly enhance the expression of CD25 and HLA-Dr activation antigens on T-cells and increased the secretion, into the medium, of IL-2 and gamma interferon. NK-cell activity was also increased by BSS and BSSG alone, but with ESF a higher activity was always found at different effector:target ratios (100:1–12:1).

HUMAN PROSTATIC-CARCINOMA CELLS MODULATE LYMPHOCYTE-PROLIFERATION BY AN IMMUNOSUPPRESSIVE ACTIVITY

We have identified an immunosuppressive activity from the conditioned medium of an androgen-independent human prostatic carcinoma cell line, JCA-1. This activity is constitutively produced by JCA-1 cells and is capable of suppressing normal human peripheral blood lymphocyte proliferation irreversibly in a dose-dependent manner. Immunosuppressive activity was semi-purified by a combination of ion-exchange chromatography and gel filtration with an apparent molecular weight of 40-55 kDa. The immunosuppressive activity was not cytolytic to lymphocytes and was sensitive to 56 degrees C, reducing agent as well as to protease digestion. Cell cycle analysis revealed that the suppressive activity did not induce apoptosis, but it prevented some G(1) lymphocytes from entering into the S phase of the cell cycle.

Proliferative Responses of Peripheral Blood Lymphocytes of Mice and Humans Stimulated with Recombinant Interleukin 2

In vitro proliferative responses of human and murine peripheral blood lymphocytes (PBL) were studied. The cells were stimulated with recombinant interleukin 2 (IL-2) or with a mixture of phorbol-13-myristate-12-acetate with either IL-2 or calcium ionophores. Proliferation of human PBL could be induced by doses of human IL-2 or phorbol that were much lower than those required to produce responses of murine cells. However, the magnitude of the responses of human PBL was definitely lower than that of the responses of murine PBL. Murine IL-2 did not induce proliferation of human PBL, but it synergized with phorbol. The responses of murine PBL reflected the previously described strain differences in responses of spleen cells that were shown to be under complex polygenic control. The proliferation of murine spleen cells could be induced with human, as well as murine IL-2, and both species of IL-2 synergized with phorbol. While no clear-cut low and high responders could be identified among normal healthy human subjects, 2 individuals produced consistently low responses to IL-2 and 1 individual displayed low responses to phorbol and calcium ionophore A23187. Available data suggest, but do not allow a firm conclusion, that proliferative responses of human PBL are under genetic control, perhaps as complex as that reported for mice.

Suppressive effect of soluble factor(s) derived from Prevotella loescheii ATCC 15930 on proliferation of human lymphocytes.

Soluble sonic extracts of Prevotella loescheii caused a dose-dependent inhibition of human peripheral blood lymphocyte proliferation by mitogen and of the proliferation of a leukemic cell line, BALL-1, when assessed by DNA synthesis (3H-thymidine incorporation). RNA (3H-uridine incorporation) and protein (3H-leucine incorporation) synthesis were similarly altered after exposure to the extract. There was no effect on cell viability as measured by either trypan blue exclusion or extracellular release of the cytoplasmic enzyme lactate dehydrogenase. Preliminary characterization indicates the suppressive factor(s) derived from P. loescheii to be a protein since it is heat-labile and trypsin-sensitive. The factor eluted in a peak on a high-pressure liquid chromatography gel filtration corresponding to a molecular weight of approximately 32,000. Since black-pigmented anaerobic rods have been implicated in the pathogenesis of periodontal disease, the data suggest that P. loescheii contributes to the disease process by suppressing lymphocyte function.

Polyunsaturated fatty acids suppress human peripheral blood lymphocyte proliferation and interleukin-2 production.

1. The effects of a variety of fatty acids on human peripheral blood lymphocyte proliferation stimulated by concanavalin A or purified protein derivative of Mycobacterium tuberculosis were studied. 2. The proliferative response to concanavalin A was inhibited by all of the polyunsaturated fatty acids tested (eicosapentaenoate, arachidonate, docosahexaenoate, linoleate and alpha-linolenate) and also by the saturated fatty acid, stearate. The greatest inhibition of proliferation (approximately 85%) was caused by eicosapentaenoate. 3. The proliferative response to the purified protein derivative of Mycobacterium tuberculosis was inhibited by all of the polyunsaturated fatty acids tested, except alpha-linolenate, and also by stearate. The greatest inhibition of proliferation (approximately 75%) was caused by eicosapentaenoate. 4. The pattern of inhibition of proliferation by fatty acids was similar to that previously reported for rat lymphocytes with one exception: oleate did not inhibit human lymphocyte proliferation. 5. The proliferation of T-lymphocytes is dependent upon their ability to synthesize and secrete the cytokine, interleukin-2. In the presence of mitogen the concentration of interleukin-2 in the culture medium increased markedly above that in the medium of non-stimulated cells. 6. All polyunsaturated fatty acids tested caused a decrease in the concentration of interleukin-2; the greatest decrease (approximately 90%) was caused by eicosapentaenoate. 7. There was a good correlation between lymphocyte proliferation in the presence of fatty acids and interleukin-2 concentration. However, stearate did not decrease the interleukin-2 concentration but did inhibit lymphocyte proliferation.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of synthetic and naturally occuring flavonoids of mitogen-induced proliferation of human peripheral-blood lymphocytes

Examination was made of the effects of 17 synthetic and naturally occurring flavonoids on human lymphocyte proliferation in the presence of concanaval in A as a mitogen. Twelve of the flavonoids examined were mono-hydroxy or methoxy derivatives. The mitogen-induced response of lymphocytes was evaluated from the extent of the incorporation of [ 3H]thymidine into cell in vitro . All the compounds showed inhibitory effects; 4.5–77.7% of [ 3H]thymidine incorporation was blocked by an 1.0 μg/m 1 concentration. The viability of lymphocytes before and after treatment, as assessed by a dye exclusion test, indicated no change, and thus the flavonoids may inhibit DNA synthesis. The flavonoids possessing 5-hydroxyl, 5-methoxyl and 6-methoxyl groups, and those with cyclohexyl instead of phenyl substituent (i.e. 2-cyclohexyl-benzopyran-4-one), showed the greatest inhibition. The inhibitory effect of any one of them was less than one half that of prednisolone, but essentially the same or somewhat exceeding that of bredinine or azathioprine. It would thus appear that the well-known anti-inflammatory effects of flavonoids may possibly arise in part from the inhibition of the proliferative response of lymphocytes.

Role of interleukin 2 on enhancement of concanavalin A-induced human peripheral blood lymphocyte proliferation by murine B cell mitogens.

Murine B cell mitogens such as bacterial lipopolysaccharide (LPS), butanol-extracted water soluble adjuvant (Bu-WSA), dextran sulfate (DS), synthetic muramyl dipeptide (MDP), and its analog MDP-Lys (L18) do not show any mitogenic ability in vitro on human peripheral blood lymphocytes or mixed cell populations of purified T and B cells obtained from the lymphocytes in an ordinary culture system. However, these mitogens are capable of enhancing the mitogenic effect of concanavalin A (Con A) in the cultures. In the presence of one of these mitogens, the activity of interleukin 2 (IL 2), but not interleukin 1, in the supernatants obtained from cultures containing Con A-stimulated T cell and B cell populations was higher than that of control cultures. The role of the newly produced IL 2 in the synergistic effect of the mitogens in human lymphocyte cell cultures was discussed.