Immunological indices in differential diagnosis of active and inactive chronic endocervicitis

A total of 132 patients were examined for chronical endocervicitis by clinical, immunological and morphological methods. According to the results obtained the patients were subdivided into several groups. The first group covered 77 females with an active clinical somatic form of the disease. The second group included 55 patients with chronical inactive endocervicitis. Controls included 25 practically healthy females. No differences were found between immunological indices of peripheral blood in groups of patients with active and inactive forms of chronic endocervicitis. Immunological tests on cervical mucus in patients with chronic endocervicitis revealed differences in the total number of leukocytes, content of living cells, functional activity of neutrophils, concentration of immunoglobulins and proinflammatory cytokines. This study may contribute to the development of immunological criteria for diagnosis of inflammatory process activity as well as a diagnostic model.     

The effect of antifungal agents on the chemoluminescence of polymorphonuclear leukocytes

The intensity of the formation of active forms of oxygen by Staphylococcus-activated polymorphonuclear leukocytes (PMNL) was registered according to the level of luminol-dependent chemiluminescence. Peritoneal PMNL of rats were shown to be already in an activated state. Activation probably occurred in the process of their penetration into the abdominal cavity. For this reason, further studies of three chloral and griseofulvin derivatives with respect to their influence on the metabolic activity of phagocytes were made on PMNL of human peripheral blood. The compounds under study were found to produce a suppressive effect, which was probably linked with their influence of the enzymatic cell systems taking part on the formation of active forms of oxygen.     

Suppressor T cells are activated in vivo in patients with multiple sclerosis coinciding with remission from acute attack

By a variety of assays, investigators (1-4) from several laboratories previously observed changes in numbers of thymus-derived suppressor lymphocytes (Ts) in patients with acute active multiple sclerosis (MS). Changes in numbers of Ts in the peripheral blood compartment of such patients closely follow disease activity. Extending these observations we now report Ts are activated in the peripheral blood of certain patients with MS. This activation occurred in vivo in the majority of patients with active disease, but only during remission after an acute episode of MS. In contrast, activated Ts were found infrequently in patients with chronic progressive or stable MS, with neurologic diseases other than MS, or healthy individuals. The suppressor activity found in peripheral blood T cells during the acute remitting stage of disease was associated with elevated numbers of lymphocytes bearing Fc receptors for IgG and/or OKT8 markers.     

Elevated anti-parasitic activity in peripheral blood monocytes and neutrophils of cattle infected with Babesia bovis

The innate immune response to bovine Babesia bovis infection in vivo has not previously been established. We used assays measuring phagocytosis and oxidative burst to investigate the immune response because they are indicative of the innate antimicrobial capacity of monocytes and neutrophils. Monocyte and neutrophil phagocytosis is thought to be non-specific in nature and so the phagocytosis of either opsonised Zymosan or Escherichia coli was used to indicate the non-specific phagocytic capacity of monocytes and neutrophils ex vivo. The kinetics of both phagocytic and oxidative burst activity in monocytes and neutrophils were followed twice weekly from pre-inoculation (day 0) through to 31 days after inoculation. Peripheral blood monocytes were found to display a pronounced oxidative burst, but a suppressed capacity to phagocytose during a primary infection. On the other hand, neutrophils exhibited an increased phagocytic capacity and reduced oxidative activity during a primary infection. These findings identified considerable antimicrobial activity evident in peripheral blood monocytes and neutrophils from cattle exposed to B. bovis as a primary exposure. This elevated antimicrobial activity was coincident with the time that parasite numbers peaked in the circulation and occurred prior to parasite clearance. These results suggest that peripheral blood monocytes and neutrophils are active mediators in the innate immune response to a primary B. bovis.     

Cytophotometry of granulocytes in chronic granulocytic leukaemia patients. Part II. Effects of chemotherapy on cell kinetics and cytochemical parameters of granulocytes

A successful course of chemotherapy (myelosan or combined regimens) results in a two to eight fold decrease of the percentage of S + G2 cells in the bone marrow and peripheral blood and in the restoration of normal morphometric and cytochemical patterns of polymorphonuclear neutrophils in the chronic phase of CGL. In those patients of the chronic phase not responding to chemotherapy and in blast crisis the proportion of S + G2 cells did not change after a course of chemotherapy. Cytostatic drugs killed cells in all phases of the cell cycle. The initial response to chemotherapy was a slight increase of the proportion of S + G2 cells in peripheral blood, apparently as a result of the block in cell cycle transition which was followed by a rapid drop of S + G2 proportion. During chemotherapy such phenomena as an appearance of tetraploid mature neutrophils and of aneuploid clones were occasionally observed.     

Production of active oxygen by neutrophils in patients with cancer of the esophagus and stomach before and after surgical interventions]

The state of the oxygen-depended antimicrobial activity of neutrophils on oncologic patients was studied. Production of active oxygen by non-activated and activated neutrophils of peripheral blood from 19 patients (the average age of 56 years, 16 males and 3 females) with esophageal cancer and cancer of the stomach and metastasis to the esophagus was recorded by chemiluminescence before and after surgical interventions (transpleural gastrectomy, Lewis type operation) on the 1st, 3rd and 7th days after the operations. The oxygen-depended antimicrobial activity of the neutrophils was investigated by their ability to produce active oxygen when incubated with opsonized zymosan and N-formyl-methionyl-leucyl-phenylalanine. Luminol chemiluminescence recorded the whole pool of active oxygen and showed the summary activity of myeloperoxidase and NADPH-oxidase, while luceginin chemiluminescence measured formation of superoxide anion radical (*O(-)2) and evaluated the activity of NADPH-oxidase. The results showed that the procedure provided estimation of the functional state of the neutrophil leukocytes and their ability to produce active oxygen. Impairment of the myeloperoxidase- and NADPH-depended production of active oxygen by non-activated and activated neutrophils observed before operations resulted in altered synthesis of active oxygen after the surgery. This is one of the leading factors of the infection development in the end. Detection of the myeloperoxidase- and NADPH-depended production of active oxygen could be of help in revealing the groups of risk among patients at the stage of the preoperative examination and serve as a prognostic factor of the development of severe infectious complications during the postoperative period.     

Suppressor cell regulation of the spontaneously induced in vitro secondary antibody response.

The cytotoxic effect of peripheral blood lymphocytes from patients with chronic active hepatitis (CAH) on Chang liver cells was studied using a chromium-51 release assay. Cytotoxic effector cell activity was unaffected or slightly enhanced after the removal of glass-adherent mononuclear cells, indicating that the major effector cell is not a classical monocyte. Lymphocytes from 12 of 18 patients with CAH were cytotoxic, and the cytotoxic cells were contained within T-cell-depleted fractions. Further studies revealed that these effector cells were K cells, not B cells. K cells were identified as Fs- and complement receptor-bearing cells without detectable surface immunoglobulin. Serial studies in CAH showed continuous cytotoxicity in contrast to transient cytotoxicity in acute viral hepatitis, suggesting that the cytotoxic activity of lymphocytes is responsible for persistence of the disease.     

Evaluation of the phagocytic activity of peripheral blood neutrophils in patients with hyperthyroidism

In 80 women with hyperthyroidism (40 with diagnosed Graves disease and 40 with hyperactive nodular goiter) the following tests related to the function of peripheral blood neutrophils have been carried out: 1. nitrotetrazolium blue (NRT) reduction test. 2. evaluation of phagocyting activity by using latex particles and living bacteria cells, and 3. determination of immunoglobulin concentration and the C3 component of the complement in blood serum. The following features were found in the patients with hyperthyroidism: 1. the elevated values of the index of spontaneous NBT reduction which return to normal following the treatment with propranolol or metizol lasting 14 days, 2. a decrease in the phagocyting activity of neutrophils occurring with stimulation of phagocytosis by both the latex particles and bacteria cells. 3. the return to normal values of the index of neutrophils phagocyting the latex particles following two-week treatment with propranolol or metizol. It was concluded that in patients with hyperthyroidism the changes in NRT reduction and phagocyting activity of peripheral blood neutrophils return to normal following the two-week treatment of these patients with both propranolol and metizol.     

Clinical evaluation of the structural characteristics of peripheral blood leukocyte DNA in patients with viral hepatitis B

The DNA structure of peripheral blood leukocytes in healthy donors and in viral hepatitis B patients was studied by the rate of the alkaline denaturation of DNA in cell lysates. An increased rate of the DNA alkaline denaturation of cell lysates was established, which was indicative of the damages in their DNA. The most pronounced damages of DNA were found in granulocytes of patients with highly active chronic hepatitis and hepatic cirrhosis, especially in cases of simultaneous delta-virus infection. The damage of leukocyte DNA reflected probably the accumulation of cells, committed to apoptosis, in peripheral blood. Apoptosis may be induced by cytotoxic lymphocytes recognizing HBV-infected cells.     

Characterization of hemoglobin-hydrolyzing acidic proteinases in human and rat neutrophils

The nature and levels of hemoglobin (Hb)-hydrolyzing acidic proteinases including cathepsin D and cathepsin E, which were most active at pH 3.5-4.0, were enzymatically and immunochemically compared between human and rat neutrophils. By subcellular fractionation and immunoprecipitation with discriminative antibodies specific for each enzyme, cathepsin D was shown to be present in the granular content fraction of both human and rat neutrophils and to account for about 35% of the total Hb-hydrolyzing activity. Cathepsin E was observed mainly in the cytoplasmic fraction of rat neutrophils from peripheral blood and peritoneal exudates and accounted for about 65% of the total activity, but it was not detected in human blood neutrophils. Immunoelectron microscopy on rat neutrophils revealed that cathepsin D was exclusively confined to lysosomes, whereas cathepsin E was localized mainly in the cytoplasmic matrix and often in the perinuclear spaces and the rough endoplasmic reticulum. The non-cathepsin D activity in human neutrophils, which represented about 65% of the total activity, appeared to be due to a serine proteinase, since it was inhibited by diisopropyl fluorophosphate and phenylmethylsulfonyl fluoride and was not inhibited by agents specific for aspartic-, cysteine-, or metallo proteinases. The enzyme(s) responsible for this activity was largely associated with the granular membranes, and a half of it could be described as an integral membrane protein on the basis of phase separation with Triton X-114 at 35 degrees C. The levels of these Hb-hydrolases in gingival crevicular fluid from human chronic inflammatory periodontitis patients were examined in order to clarify their participation in the periodontal tissue breakdown.(ABSTRACT TRUNCATED AT 250 WORDS)     

Molecular mechanisms underlying delayed apoptosis in neutrophils from multiple trauma patients with and without sepsis

Delayed neutrophil apoptosis and overshooting neutrophil activity contribute to organ dysfunction and subsequent organ failure in sepsis. Here, we investigated apoptotic signaling pathways that are involved in the inhibition of spontaneous apoptosis in neutrophils isolated from major trauma patients with uneventful outcome as well as in those with sepsis development. DNA fragmentation in peripheral blood neutrophils showed an inverse correlation with the organ dysfunction at d 10 after trauma in all patients, supporting the important role of neutrophil apoptosis regulation for patient's outcome. The expression of the antiapoptotic Bcl-2 protein members A1 and Mcl-1 were found to be diminished in the septic patients at d 5 and d 10 after trauma. This decrease was also linked to an impaired intrinsic apoptosis resistance, which has been previously shown to occur in neutrophils during systemic inflammation. In patients with sepsis development, delayed neutrophil apoptosis was found to be associated with a disturbed extrinsic pathway, as demonstrated by reduced caspase-8 activity and Bid truncation. Notably, the expression of Dad1 protein, which is involved in protein N-glycosylation, was significantly increased in septic patients at d 10 after trauma. Taken together, our data demonstrate that neutrophil apoptosis is regulated by both the intrinsic and extrinsic pathway, depending on patient's outcome. These findings might provide a molecular basis for new strategies targeting cell death pathways in apoptosis-resistant neutrophils during systemic inflammation.     

CD177~+中性粒细胞在溃疡性结肠炎患者外周血中的表达及临床意义

目的:通过对比CD177~+中性粒细胞在溃疡性结肠炎(UC)患者与正常对照者外周血中的表达差异,分析CD177~+中性粒细胞在溃疡性结肠炎发生发展中的临床意义。方法:收集30例UC患者及20例正常对照者外周血,采用流式细胞术检测中性粒细胞CD177~+中性粒细胞表达情况,对比两组CD177表达差异。结果:UC患者外周血中CD177~+中性粒细胞表达明显高于正常对照组(P 0. 01),中度活动UC外周血CD177~+中性粒细胞表达较轻度者明显增高(P 0. 05),UC患者外周血CD177~+中性粒细胞%与Mayo评分呈显著正相关(r=0. 384,P=0. 036)。结论:CD177~+中性粒细胞在UC患者外周血表达明显增高,且与疾病活动程度密切相关,能够反映UC患者临床疾病活动程度。     

Expression of the surface membrane receptors of neutrophils in myeloproliferative diseases

For 147 patients with myeloproliferative diseases, a study was made of the expression of Fc-receptors to immunoglobulins IgC, IgA, and that of FcH-receptor and receptors to C3-components of the complement in peripheral blood neutrophils. The data obtained show the lower level of neutrophils having membrane receptors in patients with acute myeloblastic leukemia and chronic granulocytic leukemia at the stage of blast transformation. A decrease in expression of membrane receptors of neutrophils was shown in patients with chronic granulocytic leukemia and benign subleukemic myelosis. The finding of a higher level of neutrophils having surface receptors revealed in patients with real polycythemia is close to the data obtained in the study of expression of membrane receptors in patients with chronic myeloproliferative diseases and healthy persons.     

Differential synthesis of 5-lipoxygenase in peripheral blood and synovial fluid neutrophils in rheumatoid arthritis

In order to investigate 5-lipoxygenase enzyme regulation in neutrophils during an inflammatory reaction, we studied 5-lipoxygenase mRNA levels, as well as de novo enzyme synthesis, in resting and activated neutrophils isolated from normal individuals and patients with rheumatoid arthritis. The approach used was to analyze these activities in resting peripheral blood neutrophils of normal individuals on the one hand and in peripheral blood and matched synovial fluid neutrophils isolated from patients with rheumatoid arthritis on the other hand. Our first observation was that resting peripheral blood neutrophils of either normal individuals or patients show detectable levels of 5-lipoxygenase mRNA and are able to synthesize the enzyme de novo. Our second observation was that inflammatory activated neutrophils from synovial fluid reveal lower 5-lipoxygenase mRNA levels and enzyme synthesis than do the patient-matched peripheral blood cells. This is in spite of the fact that, for other proteins, synovial fluid neutrophils are equally or more active than their peripheral blood counterparts. We conclude that peripheral blood neutrophils are capable of synthesizing the enzyme, thus ensuring the turnover of the protein. Furthermore, complex regulatory mechanisms appear to take place in response to inflammation as it occurs in synovial fluids of patients with rheumatoid arthritis, leading to decreased mRNA levels and enzyme synthesis. Possible mechanisms of regulation are discussed and are presently under investigation.     

Phagocytosis and peroxidase activity in neutrophils from peripheral blood of patients with malignant tumours of lung, stomach and large intestine

The phagocytic activity as revealed by latex ingestion and cytochemically demonstrated peroxidase activity of neutrophils from peripheral blood was investigated in 26 patients with pulmonary cancer, 22 patients with gastric cancer and 8 patients with tumours of large intestine and compared with results obtained in 40 healthy individuals. Statistically significant differences were found in case of both parameters between patients with pulmonary cancer and those with tumours of large intestine. The lowered phagocytic activity of neutrophils was accompanied by an enhanced peroxidase activity and vice versa. No statistically significant differences were observed, however, between the investigated groups of patients with cancer and the control group.     

Changes in leukocyte enzyme activity during experimental amyloidogenesis and in patients with amyloidosis]

The activity of hydrolytic and redox myeloperoxidase enzymes was determined in the neutrophils and lymphocytes of the peripheral blood of patients with secondary amyloidosis and also of the animals with amyloidogenesis and amyloidosis (caseine model). During the amyloidogenesis the activity of hyocrolytic enzymes and of myeloperoxidase in the neutrophils was found to decrease; this was particularly marked at the stage of the initial amyloid deposition. Changes in the enzyme activity in the animals against the background of already developed amyloidosis coincided with such in the blood cells of patients with secondary amyloidosis. The results obtained are discussed from the aspects of resorption theory.     

Hematological changes in the mice transplanted with methylcholanthrene-induced fibrosarcoma

Hematological values of the peripheral blood in the mice transplanted with methylcholanthrene-induced fibrosarcoma were examined. Values for red blood cells and platelets reduced after tumor transplantation. Values for white blood cells increased with the percentage of neutrophils being increased significantly. Phagocytic activity of neutrophils from the tumor-bearing mice was not reduced. The cultured fluid of the tumor used in the experiments possessed strong colony stimulating activity to bone marrow cells.     

Deficiency of beta-glucuronidase in neutrophils from patients with precancerous states of the larynx.

Activity of beta-glucuronidase (GR) and acid phosphatase (AP) has been determined in peripheral blood neutrophils from 24 men with precancerous states of the larynx that is leukoplakia papillomas and pachydermia by means cytochemical methods described by Hayashi et al., and Barka and Anderson, respectively. The results obtained were expressed in terms of absolute counts of enzyme-positive and enzyme-negative cells with regard to enzyme activity variation within the enzyme-positive neutrophil population; the enzyme activity index score has been calculated. The control group consisted of 20 healthy subjects of the same sex. No significant alterations were found so far as AP activity is concerned between the group studied. In contrast, activity of GR in patients with precancerous states exhibited significant lowering. The most striking feature was in almost complete absence from the blood of GR-positive neutrophils with high activity of the enzyme. Majority of these cells showed only traces of the GR activity. According to authors opinion the deficiency of GR in neutrophils of patients with precancerous lesions pertains to problem of neutrophil-mediated cytotoxic effect against mammalian tumour cells.     

Scanning electron microscope cytochemistry of normal and leukaemic leukocytes

Backscattered Electron Imaging (BEI) is a particular technique which permits to study cytochemical reactions with the Scanning Electron Microscope (SEM). The BEI data pertaining to specific enzymatic activities can be directly correlated to the surface morphology of each individual cell. Leukocytes from 5 normal individuals, 14 patients with acute nonlymphoblastic leukaemia (ANLL), 7 patients with chronic myeloid leukaemia (CML) and 3 patients with acute lymphoblastic leukaemia (ALL) were studied for myeloperoxidase activity, acid phosphatase localization, silver staining of the nuclei and phagocytosis of iron carbonyl in the BEI mode of SEM. Some normal peripheral blood leukocytes which cannot be distinguished by their surface morphology alone were satisfactorily identified with the BEI technique. Leukaemic myeloid cells can be recognized in many cases because of their positive myeloperoxidase reaction, while monocytic elements can be characterized by the presence of surface ruffles, acid phosphatase activity and active phagocytosis. The usefulness of the BEI technique in identifying different blood cell types with the SEM and its possible application to the diagnosis of certain cases of leukaemia are discussed.     

Effects of a murine mammary tumor on in vivo and in vitro hemopoiesis

The effects of an autologous transplanted mammary tumor (RIII-T3) on hemopoiesis in RIII mice are described. Tumor-bearing animals died 30 to 40 days after inoculation and displayed splenomegaly, extreme neutrophilia, and moderately increased monocyte levels in the spleen, peripheral blood, and bone marrow. The precursors of neutrophils and monocytes, granulocyte/macrophage colony-forming cells (GM-CFC) were elevated in the spleen, bone marrow, and peripheral blood. RIII-T3-conditioned medium stimulated bone marrow GM-CFC and caused the myelomonocytic cell line, WEHI-3B, to differentiate in vitro. The conditioned medium did not stimulate erythroid, megakaryocyte, or eosinophil colony formation. When conditioned medium was fractionated, two peaks of activity corresponding to GM-CSF and G-CSF were observed, suggesting that the extreme neutrophilia observed in tumor-bearing animals may result from chronic exposure of the hemopoietic system to these hemopoietic hormones.