scholarly journals IMMUNOLOGICAL REACTIVITY AND PERIPHERAL BLOOD DNA-CYTOMETRY IN CHILDHOOD ONCOLOGICAL DISEASES

2014 ◽  
Vol 12 (1-2) ◽  
pp. 65
Author(s):  
Z. R. Khamatdinova ◽  
N. F. Khisamova ◽  
R. M. Khajrullina
Keyword(s):  
Peripheral Blood ◽  
Immunological Reactivity ◽  
Dna Cytometry ◽  
Oncological Diseases

scholarly journals Monoclonal antibodies of IKO series in the three-color analysis of sub-populations of peripheral blood T-lymphocytes of oncological patients

2018 ◽  
Vol 17 (2) ◽  
pp. 47-56
Author(s):  
E. N. Zakharova ◽  
T. N. Zabotina ◽  
O. V. Korotkova ◽  
M. E. Kopyrulina ◽  
I. V. Panichenko ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Flow Cytometry ◽  
Oral Mucosa ◽  
Peripheral Blood ◽  
Specific Activity ◽  
Peripheral Blood Lymphocytes ◽  
Lymphoid Cells ◽  
Color Analysis ◽  
Oncological Patients ◽  
Oncological Diseases

Background.Clinical studies have shown that the most oncological patients have impaired immunological reactivity. Monitoring of the immune system status of patients with solid tumors includes a subpopulation analysis of immunocompetent cells. It is based on the study of expression of surface antigens and intracellular structures, and an analysis of the functional activity of immunocompetent cells. Today two, three, and even four-color reagents are used for subpopulation analysis of lymphocytes. Fluorescent conjugate with monoclonal antibodies (MAB) of the ICO series and fluorescent dyes Imd-306, Imd-506 (cyanine) and phycoerythrin (a group of phycobiliproteins) was received and characterized in N.N. Blokhin National Medical Research Center of Oncology. These conjugates have proved themselves in one and two-color analysis of subpopulations of peripheral blood lymphocytes by flow cytometry.The purposeof this study is to assess the possibility of sharing several immunofluorescent probes (IFP) on the basis of different Mab series of ICO and dyes FITC, Imd306, Imd506 and PE for three-color analysis of lymphocyte subpopulations of donors and patients with oncological diseases by flow cytometry method.Materials and methods.A panel of immunofluorescent probes (IFP) CD3FITC, CD4PE, CD8-Imd506, CD4-Imd306 was used in the work. Probes are obtained with MAB (ICO series) and fluorophores FITC, PE, Imd-306, Imd-506. The specific activity of the probes was evaluated in the immunofluorescence reaction by flow cytometry. Donor’s peripheral blood was used to optimize the conditions of the immunofluorescence reaction. Clinical approbation of the obtained sets was carried out on 2 groups of patients with oncological diseases before and after surgical treatment. The first group included 64 patients with cancer of the oral mucosa. The second group consists of 35 patients with ovarian cancer.Results. We have shown that for the three-color analysis of human lymphocytes by flow cytometry, it is recommended to use sets of labeled MAB: CD3-FITC (10 μg/ml) + CD4-Imd306 at a concentration of 5–10 μg/ml; CD3-FITC + CD4-PE at a concentration of 20 μg/ml and CD3-FITC + CD8 Imd506 at a concentration of 10 μg/ml. The final concentration of immunoglobulin in the solution of IPS is indicated. It was shown that even before the beginning of treatment in patients with cancer of the oral mucosa the subpopulation CD3+ CD4+ content was significantly lowered in the structure of T cells, and the total level of CD8+ lymphocytes significantly exceeds the parameters of the donor group. Analysis of linear CD markers of lymphoid cells in patients with ovarian cancer revealed a significant decrease in the total number of CD3+ T-lymphocytes in comparison with donors. There was no evidence of the effect of surgical treatment on the subpopulation structure of lymphoid cells.Conclusion.Combinations of fluorescent probes CD3-FITC/CD4-Imd306/CD8-Imd506 and CD3-FITC/CD4-PE/CD8-Imd506 can be used for trichromatic analysis by flow cytometry of the lymphocyte subpopulations of both healthy donors and cancer patients. Comparative studies of the specific activity of combinations of IFP based on Mab ICO series and reference commercial test systems (BD Biosciences) on lymphocytes of healthy donors and oncological patients (ovarian cancer, oral mucosa cancer) showed comparable results. The application of the obtained probes made it possible to reveal violations of the subpopulation structure of peripheral blood lymphocytes in patients with ovarian cancer and cancer of the oral mucosa, in particular, CD3 – /CD8+, CD3+/CD4+, CD3+.

scholarly journals Circulating and disseminated tumor cells as a possible prognostic factor for oncological diseases

2021 ◽  
Vol 23 (3) ◽  
pp. 490-495
Author(s):  
Andrei D. Kaprin ◽  
Svetlana V. Zatsarenko ◽  
Sergei A. Ivanov ◽  
Lyudmila Yu. Grivtsova
Keyword(s):  
Bone Marrow ◽  
Tumor Cells ◽  
Peripheral Blood ◽  
Disseminated Tumor Cells ◽  
Clinical Predictors ◽  
Modern Methods ◽  
Different Types ◽  
Oncological Diseases ◽  
New Generation

Background. Distant metastasis remains the main cause of death for patients, despite some successes in the diagnosis and treatment of solid tumors. This demonstrates the significance to study the methods for the timely detection of tumor micrometastases and to find the role of tumor cells preserved in peripheral blood or disseminated tumor cells in bone marrow after the end of the treatment. Aim. To generalize the data concerning the significance and methodology of the determining circulating tumor cells in peripheral blood and disseminated tumor cells in bone marrow in case of different types of cancer. Materials and methods. Foreign and domestic articles concerning this topic, published in eLibrary, PubMed, Medline, Scopus databases have been analyzed. Results. The literary review deals with the modern methods of determination, we have analyzed the importance of identification of circulating and disseminated tumor cells as the clinical predictors for specific nosological types of cancer. Conclusion. The enumeration of circulating and disseminated tumor cells could be a new generation of screening and evaluation of the efficacy of cancer therapy. However, it is necessary and up to date to develop the methodology of their quantitative and qualitative identification and to find the methods for evaluating functional activity.

scholarly journals IDENTIFICATION OF CELLS OBTAINED FROM BLOOD OF ONCOLOGICAL PATIENTS WITH HEMOCYTOFILTRATION

2018 ◽  
Vol 23 (2) ◽  
pp. 84-89
Author(s):  
Olga G. Grigoruk ◽  
A. F Lazarev ◽  
A. A Chimitov ◽  
T. D Khankhashanova ◽  
L. M Bazulina ◽  
...  
Keyword(s):  
Light Microscopy ◽  
Tumor Cells ◽  
Peripheral Blood ◽  
Single Cells ◽  
Technical Aspects ◽  
Circulating Cells ◽  
Oncological Patients ◽  
Oncological Diseases ◽  
Immunocytochemical Techniques

This article presents the outcomes of our own research of the detection of circulating cells in the peripheral blood of 48 patients with oncological diseases, using hemocytofiltration. The observed circulating cells (21 cases, 43.8%) have been represented by three variants: more often in the form of “bare” atypical nuclei (76.2% cases), also single conserved tumor cells (14.3%) and large shapeless single cells, probably of non-epithelial nature (9.5%). The difficulties of identifying the obtained cells in light microscopy and technical aspects of the usage of immunocytochemical techniques have been discussed.

scholarly journals EFFECT OF ANTIGEN-PRIMED DENDRITIC CELL-BASED IMMUNOTHERAPY ON ANTITUMOR CELLULAR IMMUNE RESPONSE IN PATIENTS WITH COLORECTAL CANCER

2021 ◽  
Vol 23 (4) ◽  
pp. 963-968
Author(s):  
V. V. Kurilin ◽  
E. V. Kulikova ◽  
A. V. Sokolov ◽  
Yu. A. Kozhevnikov ◽  
D. D. Blinova ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Immune Response ◽  
Dendritic Cells ◽  
Cancer Patients ◽  
Peripheral Blood ◽  
T Regulatory Cells ◽  
Mononuclear Cells ◽  
Tumor Antigens ◽  
Autologous Tumor ◽  
Oncological Diseases

The problem of treatment of oncological diseases is one of the most urgent for modern medicine. Existing treatment approaches are based on a surgical, radiation, chemotherapeutic approach, and the use of immunotherapy methods aimed at markers and / or specific antigens of tumors.Approaches based on the mechanisms of cellular and molecular regulation of a specific antitumor immune response have shown their high efficiency (for example, antibodies against HER2 in breast cancer), but these approaches have a number of side and undesirable effects that limit their application. Considering the central role of the mechanisms of recognition of tumor antigens and their presentation to cytotoxic cells in effective tumor elimination, it is important to search for and develop approaches to restore these mechanisms in cancer pathology. Because maturation, differentiation of dendritic cells and their main function are impaired in oncological diseases, scientific research is underway to obtain mature dendritic cells and restore the natural way of antigen presentation to effector cells.The work carried out limited clinical studies (13 patients with colorectal cancer), a previously developed protocol for obtaining antigen-primed dendritic cells of patients with colorectal cancer and their joint culture with autologous mononuclear cells in vitro. From the peripheral blood of cancer patients, dendritic cells primed with autologous tumor antigens (tumor cell lysate), which were co-cultured with their own mononuclear cells in the presence of immunoregulatory cytokines (IL-12 and IL-18). The resulting cell suspensions were purified from the culture medium and cytokines and used for a course of immunotherapy (weekly, 20-30 million cells intravenously, dropwise), consisting of 3-5 injections. At different periods of immunotherapy (before the start of the course of immunotherapy, 3 months and 6 months after the end of immunotherapy), immunological parameters were assessed in the peripheral blood of patients (immunogram (CD3+, CD3+CD4+, CD3+CD8+, CD19, CD16+CD56+-cells), the relative content of T-regulatory cells (CD4+CD25+FoxP3+-cells), myeloid suppressor cells (CD14+HLA-DR- cells)) and assessed the cytotoxic activity of peripheral blood mononuclear cells of patients against cells of the tumor line of human colorectal cancer (Colo-320).The data obtained showed that in cancer patients, against the background of ongoing immunotherapy, the indicator of the direct cytotoxic test significantly increases, which makes it possible to judge the effective stimulation of the antitumor cellular immune response. This is also indicated by an increase in the relative number of CD16+CD56+-cells (NK-cells) 3 months after immunotherapy. The study of immunosuppressive cells in the blood of cancer patients showed the absence of significant changes in CD14+HLA-DR- -cells and T-regulatory cells.Thus, limited clinical studies of immunotherapy of patients with colorectal cancer based on autologous dendritic cells primed with lysate of autologous tumor cells demonstrated an increase in the antitumor cytotoxic immune response. 

scholarly journals MITOGENIC ACTIVITY AND REGENERATION

2015 ◽  
Vol 7 (2) ◽  
pp. 23-39
Author(s):  
A V Pechersky ◽  
V I Pechersky ◽  
A B Smolyaninov ◽  
V N Velyaninov ◽  
S F Adylov ◽  
...  
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Peripheral Blood ◽  
Target Therapy ◽  
Buccal Epithelium ◽  
Blood Types ◽  
Oncological Diseases ◽  
The Russian Federation ◽  
Prostatic Gland ◽  
Stimulation Of

After 40 years at people decrease in a pool of pluripotent stem cells and increase of risk of development of oncological diseases is observed.To 11 patients aged from 52 till 76 years with a cancer of a kidney, bladder, prostatic gland III-IV stages of disease the chemotherapy or target therapy was carried out. To 4 patients of 60-82 years for restoration of regeneration it was carried out from 4 to 7 transfusions of mononuclear fraction of peripheral blood from young donors of 18-23 years, same-gender and blood types with recipients.In 1 month after carrying out chemotherapy or target therapy after development of a leukopenia in 11 patients the level of the bFGF increased on average by 1,74 times. At 4 patients from them the increase in level of the human VEGF-A on average by 1,25 times was observed, the 3rd of them had an increase in the human EGF on average by 1,13 times. In 3-6 months after the last transfusion of mononuclear fraction of peripheral blood the maintenance of hemopoietic cells predecessors of Cd34+ in peripheral blood increased on average by 3,25 times. At 4 patients the level of the bFGF decreased on average by 1,78 times, at 2 patients from them reduction of level of the human VEGF-A on average by 1,48 times was observed, the 3rd of them had a reduction an human EGF on average by 4,12 times. At all 4 patients in a buccal epithelium to decrease in an expression of p53 on average by 6,02 times, at the 3rd of them to decrease in an expression of Bcl-2 on average by 60,0 times.Violation of updating of tissues is the main reason for development of carcinogenesis in people is more senior than 40 years. The excess stimulation of mitotic activity at them can be reduced to normal level by means of restoration of number of a pool of pluripotent stem cells at transfusion of mononuclear fraction of peripheral blood from young donors of 18-23 years of identical with the recipient by blood types and a sex (The patent of the Russian Federation № 2350340).

The Growth of Herpesvirus mulatto in Human Fibroblast

1974 ◽  
Vol 32 ◽  
pp. 244-245
Author(s):  
Glennelle Washington ◽  
Philip P. McGrath ◽  
Peter R. Graze ◽  
Ivor Royston
Keyword(s):  
Rhesus Monkey ◽  
Microscopic Study ◽  
Electron Microscopic Study ◽  
Peripheral Blood ◽  
Human Fibroblast ◽  
Rhesus Monkeys ◽  
Human Fibroblasts ◽  
Electron Microscopic ◽  
Specific Antisera ◽  
Peripheral Blood Leucocytes

Herpes-like viruses were isolated from rhesus monkey peripheral blood leucocytes when co-cultivated with WI-38 cells. The virus was originally designated rhesus leucocyte-associated herpesvirus (LAHV) and subsequently called Herpesvirus mulatta (HVM). The original isolations were from juvenile rhesus monkeys shown to be free of antibody to rhesus cytomegalic virus. The virus could only be propagated in human or simian fibroblasts. Use of specific antisera developed from HVM showed no relationship between this virus and other herpesviruses. An electron microscopic study was undertaken to determine the morphology of Herpesvirus mulatta (HVM) in infected human fibroblasts.

A Quantitative Ultrastructural Study of Peripheral Blood Lymphocytes Containing Parallel Tubular Arrays in Epstein-Barr Virus and Cytomegalo-Virus Mononucleosis

1981 ◽  
Vol 39 ◽  
pp. 454-455
Author(s):  
C. M. Payne ◽  
P. M. Tennican
Keyword(s):  
Peripheral Blood ◽  
Lymphocyte Count ◽  
Epstein Barr Virus ◽  
Absolute Lymphocyte Count ◽  
Peripheral Blood Lymphocytes ◽  
Clinical State ◽  
Barr Virus ◽  
The Absolute ◽  
Epstein Barr ◽  
Tubular Arrays

In the normal peripheral circulation there exists a sub-population of lymphocytes which is ultrastructurally distinct. This lymphocyte is identified under the electron microscope by the presence of cytoplasmic microtubular-like inclusions called parallel tubular arrays (PTA) (Figure 1), and contains Fc-receptors for cytophilic antibody. In this study, lymphocytes containing PTA (PTA-lymphocytes) were quantitated from serial peripheral blood specimens obtained from two patients with Epstein -Barr Virus mononucleosis and two patients with cytomegalovirus mononucleosis. This data was then correlated with the clinical state of the patient.It was determined that both the percentage and absolute number of PTA- lymphocytes was highest during the acute phase of the illness. In follow-up specimens, three of the four patients' absolute lymphocyte count fell to within normal limits before the absolute PTA-lymphocyte count.In one patient who was followed for almost a year, the absolute PTA- lymphocyte count was consistently elevated (Figure 2). The estimation of absolute PTA-lymphocyte counts was determined to be valid after a morphometric analysis of the cellular areas occupied by PTA during the acute and convalescent phases of the disease revealed no statistical differences.

Neutrophil pseudoplatelets in subgingival plaque and crevicular fluid samples from periodontal diseased sites

1989 ◽  
Vol 47 ◽  
pp. 862-863 ◽  
Author(s):  
J Hanker ◽  
E.J. Burkes ◽  
G. Greco ◽  
R. Scruggs ◽  
B. Giammara
Keyword(s):  
Electron Microscopy ◽  
Bone Marrow ◽  
Peripheral Blood ◽  
Gingival Crevicular Fluid ◽  
Light And Electron Microscopy ◽  
Subgingival Plaque ◽  
Staining Reaction ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Crevicular Fluid

The mature neutrophil with a segmented nucleus (usually having 3 or 4 lobes) is generally considered to be the end-stage cell of the neutrophil series. It is usually found as such in the bone marrow and peripheral blood where it normally is the most abundant leukocyte. Neutrophils, however, must frequently leave the peripheral blood and migrate into areas of infection to combat microorganisms. It is in such areas that neutrophils were first observed to fragment to form platelet-size particles some of which have a nuclear lobe. These neutrophil pseudoplatelets (NPP) can readily be distinguished from true platelets because they stain for neutrophil myeloperoxidase. True platelets are not positive in this staining reaction because their peroxidase Is inhibited by glutaraldehyde. Neutrophil pseudoplatelets, as well as neutrophils budding to form NPP, could frequently be observed in peripheral blood or bone marrow samples of leukemia patients. They are much more prominent, however, in smears of inflammatory exudates that contain gram-negative bacteria and in gingival crevicular fluid samples from periodontal disease sites. In some of these samples macrophages ingesting, or which contained, pseudoplatelets could be observed. The myeloperoxidase in the ingested pseudoplatelets was frequently active. Despite these earlier observations we did not expect to find many NPP in subgingival plaque smears from diseased sites. They were first seen by light microscopy (Figs. 1, 3-5) in smears on coverslips stained with the PATS reaction, a variation of the PAS reaction which deposits silver for light and electron microscopy. After drying replicate PATS-stained coverslips with hexamethyldisilazane, they were sputter coated with gold and then examined by the SEI and BEI modes of scanning electron microscopy (Fig. 2). Unstained replicate coverslips were fixed, and stained for the demonstration of myeloperoxidase in budding neutrophils and NPP. Neutrophils, activated macrophages and spirochetes as well as other gram-negative bacteria were also prominent in the PATS stained samples. In replicate subgingival plaque smears stained with our procedure for granulocyte peroxidases only neutrophils, budding neutrophils or NPP were readily observed (Fig. 6).

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