Human lymphocyte activity

Human lymphocyte activation assay (HuLA) Proliferation, flu antigen-specific IgG Immunosuppression CoUinge et al. (2010)... 

An alternative to the traditional in vivo TDAR assay is the recently developed in vitro human lymphocyte activation (HuLA) assay using human peripheral blood mononuclear cells (PBMCs), which measures secondary influenza-specific responses. This assay is sensitive to, and can differentiate the responses of, a number of known immunosuppressive compounds with different mechanisms of action at concentrations within their respective therapeutic ranges. Various endpoints can be evaluated, including proliferation and flu antigen-specific antibody (IgM and lgG)-secreting cells (Collinge et al., 2010). 

Collinge, M., Cole, S.H., Schneider, P.A., Donovan, C.B., Kamperschroer, C., Kawabata, T.T. (2010). Human lymphocyte activation assay an in vitro method for predictive immunotoxicity testing. Journal of Immunotoxicology, 7, 357-366. 

J. Uberti, J.J. Lightbody, and R.M. Johnson, The effect of nucleosides and deoxycoformycin on adenosine and deoxyadenosine inhibition of human lymphocyte activation, J. Immunol. 123 ... 

Results of methyl parathion assays involving effects on chromosomes have also been contradictory. For sister chromatid exchange, Waters et al. (1982) reported a positive response in Chinese hamster ovary cells only in the presence of metabolic activation system, while methyl parathion tested positive without a metabolic activation system in Chinese hamster V79 cells (Chen et al. 1981), cultured normal human lymphoid cells (Chen et al. 1981 Gomez-Arroyo et al. 1987 Sobti et al. 1982), and Burkitt s l5miphoma cells (Chen et al. 1981). Chen et al. (1981) found a significant dose-related increase in sister chromatid exchange in both hamster and human cultured cells, but dose-related cell cycle delays were less pronounced in human cell lines than in V79 cells. Negative results were obtained for chromosomal aberrations in human lymphocytes without a metabolic activation system (Kumar et al. 1993). 

Singh S, Eehmann-Grube B, Boedde HW. 1984. Cytogenetic effects of paraoxon and methyl-parathion on cultured human lymphocytes SCE, clastogenic activity and cell cycle delay. Int Arch Occup Environ Health 54 195-200. 

The 2 -chloro and 2 -bromo congeners of either 748 (FIAC) or 758 (FMAU) are more cytotoxic than FIAC and FMAU, suggesting that these chloro and bromo nucleosides, in contrast to the 2 -fluoro compounds, are comparatively better substrates for deoxycytidine kinase of human lymphocytes than the substrates for viral-specific thymidine kinase. The disposition of the 2 -fluoro group may also be important from the biological viewpoint. It should be noted that the structural difference between RNA and DNA is at the 2 -position. The ribo type of analog (738) of FIAC is 10 times less effective in suppression of HSV replication than is FIAC. Thus Fox, and Watanabe and coworkers concluded that the 2 - up fluorine disposition and the species of the substituent at C-5 are the two important factors influencing antiviral activity. Nevertheless, the mechanism of action of 2 -deoxy-2 -fluorocytidine (737) on certain herpes viruses, including HSV-1... 

It should be remembered that some of the established antioxidants have other metabolic roles apart from free-radical scavenging. The finding of reduced antioxidant defences in diabetes, for example, may not be prima fascie evidence of increased oxidative stress, since alternative explanations may operate. For example, this may reflect a response to reduced free-radical activity as su ested by the results of a previous study (Collier et al., 1988). In the case of ascorbate, an alternative explanation has been proposed by Davis etal. (1983), who demonstrated competitive inhibition of ascorbate uptake by glucose into human lymphocytes. This view is supported by the similar molecular structure of glucose and ascorbic acid (see Fig. 12.4) and by a report of an inverse relationship between glycaemic control and ascorbate concentrations in experimental diabetes in rats. Other investigators, however, have not demonstrated this relationship (Som etal., 1981 Sinclair etal., 1991). 

Schinazi RF, Chu CK, Peck A, McMillan A, Mathis R, Cannon D, Jeong L-S, Beach JW, Choi W-B, Yeola S, Liotta DC. Activities of the four optical isomers of 2, 3 -dideoxy-3 -thiacytidine (BCH-189) against human immunodeficiency virus type 1 in human lymphocytes. Antimicrob Agents Chemo-ther 1992 36 672-676. 

Investigations of the cellular effects of radiofrequency radiation provide evidence of damage to various types of avian and mammalian cells. These effects involve radiofrequency interactions with cell membranes, especially the plasma membrane. Effects include alterations in membrane cation transport, Na+/K+-ATPase activity, protein kinase activity, neutrophil precursor membrane receptors, firing rates and resting potentials of neurons, brain cell metabolism, DNA and RNA synthesis in glioma cells, and mitogenic effects on human lymphocytes (Cleary 1990). 

No information on genotoxicity in humans was located. In vitro, a cytogenic assay with human lymphocytes was negative (Collins et al. 1995). Vapor concentrations ranged from 5% to 100% volume per volume (v/v), and the incubation period was 3 h in both the presence and absence of metabolic activation. 

Perocco P, Bolognesi S, Alberghini W. 1983. Toxic activity of seventeen industrial solvents and halogenated compounds on human lymphocytes cultured in vitro. Toxicol Lett 16 69-75. 

An alternative method for removing Zn from the active site of zinc proteins is to use aromatic nitroso compounds such as 3-nitrosoben-zamide and 6-nitroso-l,2-benzopyrone (382). These agents can oxidize Zn-bound cysteine S and can inhibit HIV-1 infection in human lymphocytes. They also eject zinc from isolated HIV-1 nucleocapsid zinc-fingers and from intact HIV-1 virions. 

When metabolic activation is used, S9 mix should not exceed 1-10 percent of the culture medium by volume. It has been shown that the S9 mix is clastogenic in CHO cells and mouse lymphoma cells (Cifone et al., 1987 Kirkland et al., 1989) but not in human lymphocytes, where blood components can inactivate active oxygen species which could cause chromosome damage. When S9 mix from animals treated with other enzyme-inducing agents such as phenobarbitone/beta-naphtho-flavone, is used, clastogenesis may be minimized (Kirkland et al., 1989). 

Kirkland D.J., Marshall R.R., McEnaney, S., Bidgwood, J., Rutter, A. and Mulliheux, S. (1989). Aroclor-1254 induced rat liver S-9 causes chromosome aberrations in CHO cells but not in human lymphocytes, a role for active oxygen Mutat. Res. 214 115-122. 

Morimoto K, Wolff S, Koizumi A. 1983. Induction of sister-chromatid exchanges in human lymphocytes by microsomal activation of benzene metabolites. Mutat Res 119 355-360. 

Macon-Lemaitre L. Triebel F The negative regulatory function of the lymphocyte-activation gene-3 co-receptor (CD223) on human T cells. Immunology 2005 115 170-178. 

Panossian A, Davtyan T, Gukassyan N, Gukasova G, Mamikonyan G, Gabrielian E, Wikman G. (2002) Effect of andrographolide and Kan Jang-fixed combination of extract SHA-10 and extract SHE-3 on proliferation of human lymphocytes, production of cytokines and immune activation markers in the whole blood cells culture. Phytomedicine 9 598-605. 

Mammalian cells in vitro are exposed to the test chemical with and without an exogenous mammalian metaboUc activation system and cultured for two rounds of replication in bromodeox3Uiridine (BrdU) containing medium. After treatment with a spindle inhibitor (e.g., colchicine) to accumulate cells in a metaphase-Uke stage of mitosis (c-metaphase), cells are harvested, stained, and metaphase cells analyzed for SCEs. Primary cultures (e.g., human lymphocytes) or established cell lines (e.g., Chinese hamster ovary or lung cells) may be used in the assay. At least three adequately spaced concentrations of the test substance should be used. 

Mutagenesis Entacapone was mutagenic and clastogenic in the in vitro mouse lymphoma/thymidine kinase assay in the presence and absence of metabolic activation, and was clastogenic in cultured human lymphocytes in the presence of metabolic activation. 

Neron S, Nadeau PJ, Darveau A et al (2011) Tuning of CD40-CD154 interactions in human B-lymphocyte activation a broad array of in vitro models for a complex in vivo situation. Arch Immunol Ther Exp (Warsz) 59 25-40... 

B17. Bussing, A., Vervecken, W, Wagner, M., Wagner, B., Pfuller, U., and Schietzel, M., Expression of mitochondrial Apo2.7 molecules and caspase-3 activation in human lymphocytes treated with the ribosome-inhibiting mistletoe lectins and the cell membrane permeabilizing viscotoxins. Cytometry 37, 133-139 (1999). 

Anti-lymphocyte globulin (ALG) has been prepared as an highly purified solution of y-globulins with antilymphocyte activity by immunizing horses with human lymphocytes. It activates complement-mediated destruction of lymphocytes and thus decreases cellular immunity with only a limited effect on humoral immunity. Anti-lymphocyte globulin suppresses delayed type hypersensitivity reactions. It is used for the prevention and treatment of rejection episodes of transplanted organs. It also has some indication for the management of idiopathic aplastic anemia. Adverse effects include pain at the site of injection, erythema, serum sickness and rarely anaphylactic shock and thrombocytopenia. 

Arrouijal FZ, Marzin D, Hildebrand HF, et al. 1992. Differences in genotoxic activity of -Ni3S2 on human lymphocytes from nickel-hypersensitized and nickel-unsensitized donors. Mutagenesis 7(3) 183-187. 

Chromosome aberration induced. Lyo-philized extract of the roasted seed, in cell culture at a concentration of 3.9 mg/mL, was active on human lymphocytes. Caffeinated and decaffeinated coffees without S9 mix was tested. The extract produced weak activity with S9 mix . Extract of the roasted seed, in cell culture at variable concentrations, was active on human lymphocytes. Metabolic activation reduced the effect . 

Fresh root juice, in the drinking water of rats at a dose of 300 mL animal, was active on Chinese hamster ovary cells. The plasma from the rats treated with carrot juice and cyclophoshamide reduced sister chromatid exchange in DNA-repair-deficient and in normal human lymphocytes in the Chinese hamster ovary cells when compared to cyclophoshamide treated only . 

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