Combined Action of PGRPs-Hsp70 Cytotoxic Complex with Paclitaxel Improves Outcomes of Melanoma Treatment in Mice.

We studied the effect of PGRPs-Hsp70 cytotoxic complex that is analogous to natural complex secreted by cytotoxic lymphocytes and the antitumor drug paclitaxel on the development of M3 melanoma in DBA mice. Significant inhibition of tumor growth was observed in all experimental groups by days 20 and 35 of observation; paclitaxel monotherapy was less effective than administration of PGRPs-Hsp70 cytotoxic complex and its combination with paclitaxel. Pairwise comparison of Kaplan-Meier curves showed that survival was maximum in the group receiving combined therapy with PGRPs-Hsp70 cytotoxic complex and paclitaxel in comparison with groups receiving monotherapy.

The role of S100A4 protein in anticancer cytotoxicity: its presence is required on the surface of CDCDPGRPsS100A4 lymphocyte and undesirable on the surface of target cells.

S100A4 is a Ca-binding protein that performs an important role in metastasis. It is also known for its antitumor functions. S100A4 is expressed by a specialized subset of CDCD lymphocytes and is present on those cell's membranes along with peptidoglycan recognition proteins (PGRPs).

Data of self-made Taq DNA polymerase prepared for screening purposes.

DNA analysis is a key procedure in genetic engineering. Nowadays the analysis is often done by PCR with Taq DNA polymerase. Although the last enzyme price is quite low, demand for numerous analyses results in much money expenditure which are not affordable for many laboratories.

Assessment of the Status of Effector and Regulatory Components of Immune System in Chelyabinsk Region Residents Exposed to Radiation Due to Residence in the Area Contaminated as a Result of the Radiation Accident at Mayak PA and in Their Offspring.

Immune status was studied in the framework of the current work and the results of the analysis of concentration of 26 characteristic parameters of innate and acquired immunity in 140 individuals from 56 trios (fathers, mothers and their Ist generation offspring that were included in 2 groups) are presented. Fathers and mothers of the children under study in the main groip Were exposed due to a long-term residence (from childhood to maturity) in the areas of Chelyabinsk region contaminated as a result of the-accident at Mayak PA (contamination included long-lived isotopes - ⁹⁰Sr and, to a smaller extent, ¹³⁷Cs and ²³⁹Pu) and then migrated into Ozyorsk prior to the conception of their children (75 individuals, 33 family trios). Comparison group (control) included parents and their offspring who are Ozyorsk residents never residing in the areas contaminated by radionuclides (65 individuals, 23 family trios).

A new role for PGRP-S (Tag7) in immune defense: lymphocyte migration is induced by a chemoattractant complex of Tag7 with Mts1.

PGRP-S (Tag7) is an innate immunity protein involved in the antimicrobial defense systems, both in insects and in mammals. We have previously shown that Tag7 specifically interacts with several proteins, including Hsp70 and the calcium binding protein S100A4 (Mts1), providing a number of novel cellular functions. Here we show that Tag7-Mts1 complex causes chemotactic migration of lymphocytes, with NK cells being a preferred target.

[The substantiation for the application of high-intensity laser therapy for the treatment of the patients presenting with gonarthrosis].

Unlabelled: The objective of the present study was to evaluate the influence of high-intensity laser irradiation on the state of microcirculation in the patients presenting with gonarthrosis.

Material And Methods: It included 40 patients with this condition at the ag from 40 to 75 years who were randomly allocated to two groups. Group 1 was comprised of 20 patients treated by high-intensity laser irradiation (HILl), the patients of group 2 (n = 20) underwent the placebo treatment.

Extracellular HspBP1 inhibits formation of a cytotoxic Tag7-Hsp70 complex in vitro and in human serum.

Tag7 (PGRP-S) was described as an innate immunity protein. Earlier we have shown that Tag7 forms with Hsp70 a stable complex with cytotoxic and antitumor activity. The same complex is formed in and secreted by cytotoxic T-lymphocytes.

The heat shock-binding protein (HspBP1) protects cells against the cytotoxic action of the Tag7-Hsp70 complex.

Heat shock-binding protein HspBP1 is a member of the Hsp70 co-chaperone family. The interaction between HspBP1 and the ATPase domain of the major heat shock protein Hsp70 up-regulates nucleotide exchange and reduces the affinity between Hsp70 and the peptide in its peptide-binding site. Previously we have shown that Tag7 (also known as peptidoglycan recognition protein PGRP-S), an innate immunity protein, interacts with Hsp70 to form a stable Tag7-Hsp70 complex with cytotoxic activity against some tumor cell lines.

Opposite roles of metastasin (S100A4) in two potentially tumoricidal mechanisms involving human lymphocyte protein Tag7 and Hsp70.

We compare the physical and functional interactions between three widespread multifunctional proteins [metastasin (Mts1/S100A4), innate immunity-related Tag7/PGRP-S, and Hsp70] in two experimental models relevant to host-tumor relationships on humoral and cellular levels. (i) Tag7 and Hsp70 in solution or in a lymphocyte make a stable binary complex that is highly cytotoxic for some tumor cells. Here, we show that Mts1 prevents Tag7.

Significance of methods for purification of oligodeoxyribonucleotide probes for the efficiency of gene diagnosis by real-time PCR.

Analysis of the use of real-time PCR with fluorescent registration of results for gene diagnosis of infectious diseases showed that the sensitivity and reliability of quantitative evaluation of DNA targets directly depended on the method of purification of oligonucleotide probes. Chromatographic behavior of synthetic probes carrying various fluorophores and fluorescence quenchers was analyzed. Approaches to optimization of purification methods are proposed enabling elimination of previously undetectable admixtures.

Comparative analysis of secretion of S100A4 metastatic marker by immune and tumor cells.

S100A4 protein is present in low concentrations (2.1-15.7 ng/10(6) cells) in lymphocyte and neutrophil culture medium.

Interactions and possible functional characteristics of Tag7-S100A4 protein complex.

Peptidoglycane-recognizing protein Tag7 formed a complex with S100A4 (a representative of S100 protein family), the apparent dissociation constants in the absence and presence of Ca2+ were 2 x l0(-8) M and 10(-9) M, respectively. Analysis of fluorescence spectra of hydrophobic fluorescent probe 2-toluidinyl naphthalene-6-sulfonate in the presence of S100A4 and Tag7 proteins showed that extensive area or several sites are involved into the complex formation between these proteins. The formation of Tag7-S100A4 complex had virtually no effect on the role of S100A4 in the regulation of intracellular Ca2+ metabolism.

Cytotoxic T lymphocytes carrying a pattern recognition protein Tag7 can detect evasive, HLA-negative but Hsp70-exposing tumor cells, thereby ensuring FasL/Fas-mediated contact killing.

Within the broad problem of host immune surveillance versus tumor immune evasion, a most intriguing question is how the cellular immunity can cope with cancerous cells that have gotten rid of the classical antigen-presenting machinery. One such option stems from (1) the fact that HLA loss is often attended with expression of Hsp70 on the tumor cell surface, and (2) our findings that human lymphocytes express a protein Tag7 (also known as PGRP-S) capable of tight and specific interaction with cognate Hsp70. Here we show that a subpopulation of human CD4(+)CD25(+) lymphocytes, obtained either in culture as lymphokine-activated killers or directly from healthy donors, carry Tag7 and FasL on their surface and can indeed kill the HLA-negative tumor-derived cells K562 and MOLT-4 that expose Hsp70 and Fas.

Peptidoglycan recognition protein tag7 forms a cytotoxic complex with heat shock protein 70 in solution and in lymphocytes.

The peptidoglycan recognition protein Tag7 is shown to form a stable 1:1 complex with the major stress protein Hsp70. Neither protein is cytotoxic by itself, but their complex induces apoptotic death in several tumor-derived cell lines even at subnanomolar concentrations. The minimal part of Hsp70 needed to evoke cytotoxicity is residues 450-463 of its peptide-binding domain, but full cytotoxicity requires its ATPase activity; remarkably, Tag7 liberated from the complex at high ATP is not cytotoxic.

Mechanisms of hemopoiesis in athymic mice.

Cell elements of peripheral blood and bone marrow and splenic hemopoiesis were studied in athymic BALB/c mice (nude or thymectomized animals). In athymic animals lympho- and erythropoiesis were suppressed, while granulocyto- and monocytopoiesis were activated. These changes were more pronounced in nude mice.

Activity of sympathoadrenal system and myelokaryocyte death during aging in AKR/JY mice.

Accelerated bone marrow cell death and activation of the sympathoadrenal system were observed during aging of highly leukemic 2-7-month-old AKR/JY mice compared to that in (CBA/CaLacxAKR/JY)F1 strain. Close correlation was revealed between activity of the sympathoadrenal system and necrotic and apoptotic forms of cell death. This can promote tumor process, because maximum changes in hemopoietic cells occur during advanced stage of the disease.

Apoptotic and necrotic cytolytic processes induced by membrane associated proteins of LAK cells.

Cytolytic processes induced by membrane-associated proteins of human lymphokine-activated killer (LAK) cells with different phenotypes (CD3+, CD16-, CD8+ and CD16+, CD8+, CD3-) were studied using L929 and K562 types of target cells. Independently of the phenotype of effector cells and the type of target cells, total fractions of membrane proteins induced several different cytolytic processes occurring with different rates and involving different mechanisms of genome fragmentation. The membrane fraction induced, irrespective of the phenotype of LAK cells, mostly apoptotic processes in the L929 line.

Different pathways of the release of cytotoxic proteins in LAK cells.

Human LAK cells were shown to release cytotoxic proteins by both Ca(2+)-dependent and Ca(2+)-independent mechanisms. CD3+ CD8+ CD16- and CD16+ CD8+ CD3- LAK cells were co-incubated with target cells in the presence of 4 mM EGTA. Although EGTA inhibited the exocytosis of cytolytic granules, supernatants obtained were cytotoxic for target cells.

Effect of antibodies to granular perforin and calmodulin on the activity of natural killer cytotoxic factor.

It was previously established that the cell death induced by natural killer cytotoxic factor (NKCF) is a complex calcium-dependent two-stage process. In the initial stage, during the first 30 min of incubation of NKCF with target cells, pore formation on the surface of the target is observed, together with temporary membrane damage. The second stage cytolytic activity occurs after a 3 h latent period, reaches maximum at 24 h, and leads to cell death.