Tissue contexture determines the pattern and density of tumor-infiltrating immune cells in HPV-associated squamous cell carcinomas of oropharynx and uterine cervix.

The profile of the antitumor immune response is an important factor determining patient clinical outcome. However, the influence of the tissue contexture on the composition of the tumor microenvironments of virally induced tumors is not clearly understood. Therefore, we analyzed the immune landscape of two HPV-associated malignancies: oropharyngeal squamous cell carcinoma (OPSCC) and squamous cell carcinoma of uterine cervix (CESC). We employed multiplex immunohistochemistry and immunofluorescence to evaluate the density and spatial distribution of immune cells in retrospective cohorts of OPSCC and CESC patients. This approach was complemented by transcriptomic analysis of purified primary tumor cells and in silico analysis of publicly available RNA sequencing data. Transcriptomic analysis showed similar immune profiles in OPSCC and CESC samples. Interestingly, immunostaining of OPSCC tissues revealed high densities of immune cells in both tumor stroma and tumor epithelium, whereas CESC samples were mainly characterized by the lack of immune cells in the tumor epithelium. However, in contrast to other immune cell populations, polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs) were abundant in both segments of CESC samples and CESC-derived tumor cells expressed markedly higher levels of the PMN-MDSC chemoattractants CXCL1, CXCL5, and CXCL6 than OPSCC tumor cells. Taken together, despite their having the same etiologic agent, the immune infiltration pattern significantly differs between OPSCC and CESC, with a noticeable shift toward prominent MDSC infiltration in the latter. Our data thus present a rationale for a diverse approach to targeted therapy in patients with HPV-associated tumors of different tissue origins.

Ways to reduce the labor intensiveness of applying the method of optical estimation of microbial cell concentration in suspension.

The labor intensity (in hours) of the optical method of microbial cell counting in suspension compared to the method of microbial cell counting using a Goryaev chamber is evaluated. The relevance of assessing the production labor intensity of microbial cell counting methods in suspension is related to the need to use them in many studies. Often the commonly used methods are too labour-intensive, time-consuming, or require expensive equipment. A comparative experiment was carried out with our previously developed "Method for optical estimation of microbial cell concentration in suspension" (Priority certificate No. 2016141859 dated 25.10.2016) and the method of microbial cell counting using a Goryaev chamber. Production labor intensity of the measurements performed was calculated in hours according to the formula: Tp=Tt+Tob, where Tp is production labour input, Tt is technological labour input, Tob is maintenance labour input. Technological labour input of measurements with use of Goryaev's chamber made up 32,18 ± 0,95, whereas with optical method - 1,03±0,06 (reliability of differences at p<0,01) at amount of measurements n = 100. Labour input of service at optical method 0,24 ± 0,03, at application of method with use of Goryaev chamber 0,15±0,01 hours. Labour input of measurements of concentration of microbial cells in suspension at application of method of measurement with Goryaev chamber remains (p<0,01) higher than at an optical method of estimation, 32,33±0,96 and 1,27±0,05 hours accordingly. When using the optical method of concentration estimation in the suspension it is necessary to carry out not a small amount of necessary mathematical calculations, which in the future, probably, corrected by creating a special program for a personal computer. The labour input of results obtained by measuring by optical evaluation of the concentration of microbial cells in suspension is lower than that obtained by using a measurement method using a Goryaev chamber. Taking into consideration that its implementation does not require purchase of special equipment as in turbidimetry, its cost-effectiveness compared to existing ones is obvious.

Tumor-initiating cells of breast and prostate origin show alterations in the expression of genes related to iron metabolism.

The importance of iron in the growth and progression of tumors has been widely documented. In this report, we show that tumor-initiating cells (TICs), represented by spheres derived from the MCF7 cell line, exhibit higher intracellular labile iron pool, mitochondrial iron accumulation and are more susceptible to iron chelation. TICs also show activation of the IRP/IRE system, leading to higher iron uptake and decrease in iron storage, suggesting that level of properly assembled cytosolic iron-sulfur clusters (FeS) is reduced. This finding is confirmed by lower enzymatic activity of aconitase and FeS cluster biogenesis enzymes, as well as lower levels of reduced glutathione, implying reduced FeS clusters synthesis/utilization in TICs. Importantly, we have identified specific gene signature related to iron metabolism consisting of genes regulating iron uptake, mitochondrial FeS cluster biogenesis and hypoxic response (ABCB10, ACO1, CYBRD1, EPAS1, GLRX5, HEPH, HFE, IREB2, QSOX1 and TFRC). Principal component analysis based on this signature is able to distinguish TICs from cancer cells in vitro and also Leukemia-initiating cells (LICs) from non-LICs in the mouse model of acute promyelocytic leukemia (APL). Majority of the described changes were also recapitulated in an alternative model represented by MCF7 cells resistant to tamoxifen (TAMR) that exhibit features of TICs. Our findings point to the critical importance of redox balance and iron metabolism-related genes and proteins in the context of cancer and TICs that could be potentially used for cancer diagnostics or therapy.