scholarly journals Defective mononuclear leukocyte chemotaxis in the Chediak-Higashi syndrome of humans, mink, and cattle

Blood ◽  
1975 ◽  
Vol 45 (6) ◽  
pp. 863-870 ◽  
Author(s):  
JI Gallin ◽  
JA Klimerman ◽  
GA Padgett ◽  
SM Wolff
Keyword(s):  
Capillary Tube ◽  
Autologous Serum ◽  
Mononuclear Leukocytes ◽  
Boyden Chamber ◽  
Mononuclear Leukocyte ◽  
Leukocyte Chemotaxis ◽  
Filter Assay ◽  
Chediak Higashi Syndrome ◽  
Micropore Filter

Chemotaxis of mononuclear leukocytes from humans, mink, and cattle was evaluated in vitro using a morphologic Boyden chamber technique and a new 51-Cr-labeled mononuclear radioassay with a double micropore filter system. Significantly decreased mononuclear leukocyte chemotactic response were noted when human, mink, or cattle Chediak-Higashi cells were tested using autologous serum or endotoxin-activated autolotous serum. A similar Chediak-Higashi mononuclear leukocyte defect was noted in humans when kallikrein or dialyzable transfer factor were used as the chemotactic stimulus. Studies using smaller pore filters in the chemotactic chamber exaggerated the chemotactic defect. Serum from Chediak-Higashi subjects had normal chemotactic activity. Additional studies on the spontaneous (random) locomotion of Chediak-Higashi mononuclear leukocytes revealed normal results when a capillary tube assay system was used, but abnormal results were obtained when a Boyden chamber micropore filter assay was used, demonstrating fundamental differences in these two assays of random locomotion. It is clear from these studies that defective mononuclear leukocyte chemotaxis is another feature of the imparied host defenses in the Chediak-Higashi syndrome that may contribute to the marked susceptibility to pyogenic infections so characteristic of this dease.

scholarly journals Defective mononuclear leukocyte chemotaxis in the Chediak-Higashi syndrome of humans, mink, and cattle

Blood ◽  
1975 ◽  
Vol 45 (6) ◽  
pp. 863-870 ◽  
Author(s):  
JI Gallin ◽  
JA Klimerman ◽  
GA Padgett ◽  
SM Wolff
Keyword(s):  
Capillary Tube ◽  
Autologous Serum ◽  
Mononuclear Leukocytes ◽  
Boyden Chamber ◽  
Mononuclear Leukocyte ◽  
Leukocyte Chemotaxis ◽  
Filter Assay ◽  
Chediak Higashi Syndrome ◽  
Micropore Filter

Abstract Chemotaxis of mononuclear leukocytes from humans, mink, and cattle was evaluated in vitro using a morphologic Boyden chamber technique and a new 51-Cr-labeled mononuclear radioassay with a double micropore filter system. Significantly decreased mononuclear leukocyte chemotactic response were noted when human, mink, or cattle Chediak-Higashi cells were tested using autologous serum or endotoxin-activated autolotous serum. A similar Chediak-Higashi mononuclear leukocyte defect was noted in humans when kallikrein or dialyzable transfer factor were used as the chemotactic stimulus. Studies using smaller pore filters in the chemotactic chamber exaggerated the chemotactic defect. Serum from Chediak-Higashi subjects had normal chemotactic activity. Additional studies on the spontaneous (random) locomotion of Chediak-Higashi mononuclear leukocytes revealed normal results when a capillary tube assay system was used, but abnormal results were obtained when a Boyden chamber micropore filter assay was used, demonstrating fundamental differences in these two assays of random locomotion. It is clear from these studies that defective mononuclear leukocyte chemotaxis is another feature of the imparied host defenses in the Chediak-Higashi syndrome that may contribute to the marked susceptibility to pyogenic infections so characteristic of this dease.

scholarly journals Dengue viruses and mononuclear phagocytes. II. Identity of blood and tissue leukocytes supporting in vitro infection

1977 ◽  
Vol 146 (1) ◽  
pp. 218-229 ◽  
Author(s):  
SB Halstead ◽  
EJ O'Rourke ◽  
AC Allison
Keyword(s):  
Bone Marrow ◽  
Dengue Infection ◽  
Neutralizing Antibody ◽  
Mononuclear Phagocyte ◽  
Mononuclear Phagocytes ◽  
Autologous Serum ◽  
Mononuclear Leukocytes ◽  
Direct Immunofluorescence ◽  
Blood Leukocytes

Studies were made on the identity of human and monkey mononuclear leukocytes permissive to antibody-enhanced dengue 2 virus (D2V) infection. In cultures of peripheral blood leukocytes (PBL) inoculated immediately after separation, it was concluded that only mononuclear phagocytes support dengue infection. This is based upon observations that D2V-permissive cells were resistant to 1,200 rads, were both plastic adherent and nonadherent, were removed when passed through nylon wool columns in 10 percent fetal bovine serum or 100 percent autologous serum, and were destroyed by incubation with 100 μg/ml particulate silica. On direct immunofluorescence staining, perinuclear dengue antigen was visualized at 24 h, becoming maximal at 60 h. Antigen-containing cells had ample cytoplasm, ruffled cytoplasmic membrane, and 73 percent were actively phagocytic. As further evidence of the infection of mononuclear phagocytes, antibody-enhanced D2V replication was observed in bone marrow cultures from five of five rhesus monkeys, but not in cell cultures of spleen, thymus, or lymph nodes prepared from the same animals. It is hypothesized that dengue virus complexed with non-neutralizing antibody is internalized by immune phagocytosis in a mononuclear phagocyte with a defective virus-destroying mechanism. Dengue permissiveness may depend upon cellular immaturity since bone marrow leukocytes could be infected even when held for 4 days before infection while PBL held for this time decreased in permissiveness. In vitro antibody-dependent infection of mononuclear phagocytes should prove useful as a model for study of immunopathologic mechanisms in human dengue.

scholarly journals Polymorphonulcear leukocyte chemotaxis toward oxidized lipid components of cell membranes.

1975 ◽  
Vol 141 (6) ◽  
pp. 1437-1441 ◽  
Author(s):  
S R Turner ◽  
J A Campbell ◽  
W S Lynn
Keyword(s):  
Arachidonic Acid ◽  
Polymorphonuclear Leukocyte ◽  
Cell Membranes ◽  
Plasma Membranes ◽  
Chemotactic Activity ◽  
Assay System ◽  
Leukocyte Chemotaxis ◽  
Filter Assay ◽  
Micropore Filter

Polymorphonuclear leukocyte chemotaxis has been elicited by oxidized arachidonic acid and other oxidized polyenoic lipids in the Boyden micropore filter assay system. This chemotactic activity was observed in the absence of serum and chemotactic proteins. The esterfied arachidonic acid present in plasma membranes may be a precursor of chemotactic messages as well as prostaglandins in vivo.

scholarly journals Granulocyte Chemotaxis in the Chediak-Higashi Syndrome of Mink

Blood ◽  
1972 ◽  
Vol 39 (5) ◽  
pp. 644-649 ◽  
Author(s):  
Robert A. Clark ◽  
Harry A. Kimball ◽  
George A. Padgett
Keyword(s):  
Chemotactic Response ◽  
In Vitro Studies ◽  
Susceptibility To Infection ◽  
Inflammatory Site ◽  
Chemotactic Factors ◽  
Leukocyte Chemotaxis ◽  
Chediak Higashi Syndrome ◽  
Increased Susceptibility

Abstract Studies of granulocyte chemotaxis were performed in mink with the Chediak-Higashi syndrome. In vivo migration of leukocytes to an inflammatory site was reduced in the affected animals. In vitro studies documented a consistent early impairment in the chemotactic response of Chediak-Higashi mink granulocytes. Serum from Chediak-Higashi mink generated normal amounts of chemotactic factors. The cellular defect in leukocyte chemotaxis in mink is comparable to that observed in humans with the Chediak-Higashi syndrome and may contribute to the increased susceptibility to infection in this disease.

scholarly journals Tomatine Displays Antitumor Potential in In Vitro Models of Metastatic Melanoma

2020 ◽  
Vol 21 (15) ◽  
pp. 5243 ◽  
Author(s):  
Simona Serratì ◽  
Letizia Porcelli ◽  
Stefania Guida ◽  
Anna Ferretta ◽  
Rosa Maria Iacobazzi ◽  
...  
Keyword(s):  
Metastatic Melanoma ◽  
Cell Lines ◽  
Cell Invasion ◽  
Capillary Tube ◽  
Tube Formation ◽  
Boyden Chamber ◽  
Wild Type ◽  
Antitumor Potential ◽  
Vegf Release

There is a growing interest in the cytotoxic effects of bioactive glycoalkaloids, such as α-tomatine on tumor cells. Here, for the first time, we determine the antitumor potential of tomatine, a mixture of α-tomatine and dehydrotomatine, in metastatic melanoma (MM) cell lines harboring different BRAF and MC1R variants. We performed cytotoxicity experiments and annexin-V/propidium iodide staining to assess the apoptotic/necrotic status of the cells. ER stress and autophagy markers were revealed by Western Blot, whereas antiangiogenic and vascular-disrupting effects were evaluated through a capillary tube formation assay on matrigel and by ELISA kit for VEGF release determination. Cell invasion was determined by a Boyden chamber matrigel assay. Tomatine reduced 50% of cell viability and induced a concentration-dependent increase of apoptotic cells in the range of 0.5–1 μM in terms of α-tomatine. The extent of apoptosis was more than two-fold higher in V600BRAF-D184H/D184H MC1R cells than in BRAF wild-type cells and V600BRAF-MC1R wild-type cell lines. Additionally, tomatine increased the LC3I/II autophagy marker, p-eIF2α, and p-Erk1/2 levels in BRAF wild-type cells. Notably, tomatine strongly reduced cell invasion and melanoma-dependent angiogenesis by reducing VEGF release and tumor-stimulating effects on capillary tube formation. Collectively, our findings support tomatine as a potential antitumor agent in MM.

scholarly journals Sex-dependent dysregulation of human neutrophil responses by bisphenol A

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Wioletta Ratajczak-Wrona ◽  
Marzena Garley ◽  
Malgorzata Rusak ◽  
Karolina Nowak ◽  
Jan Czerniecki ◽  
...  
Keyword(s):  
Nadph Oxidase ◽  
Bisphenol A ◽  
Total Concentration ◽  
Nicotinamide Adenine Dinucleotide Phosphate ◽  
Human Neutrophils ◽  
Boyden Chamber ◽  
No Production ◽  
Pathogen Elimination ◽  
Latex Beads

Abstract Background In the present study, we aimed to investigate selected functions of human neutrophils exposed to bisphenol A (BPA) under in vitro conditions. As BPA is classified among xenoestrogens, we compared its action and effects with those of 17β-estradiol (E2). Methods Chemotaxis of neutrophils was examined using the Boyden chamber. Their phagocytosis and nicotinamide adenine dinucleotide phosphate hydrogen (NADPH) oxidase activity were assessed via Park’s method with latex beads and Park’s test with nitroblue tetrazolium. To assess the total concentration of nitric oxide (NO), the Griess reaction was utilized. Flow cytometry was used to assess the expression of cluster of differentiation (CD) antigens. The formation of neutrophil extracellular traps (NETs) was analyzed using a microscope (IN Cell Analyzer 2200 system). Expression of the investigated proteins was determined using Western blot. Results The analysis of results obtained for both sexes demonstrated that after exposure to BPA, the chemotactic capacity of neutrophils was reduced. In the presence of BPA, the phagocytic activity was found to be elevated in the cells obtained from women and reduced in the cells from men. Following exposure to BPA, the percentage of neutrophils with CD14 and CD284 (TLR4) expression, as well as the percentage of cells forming NETs, was increased in the cells from both sexes. The stimulatory role of BPA and E2 in the activation of NADPH oxidase was observed only in female cells. On the other hand, no influence of E2 on the expression of CD14 and CD284, chemotaxis, phagocytosis, and the amount of NET-positive neutrophils was found for both sexes. The study further showed that BPA intensified NO production and iNOS expression in the cells of both sexes. In addition, intensified expression of all tested PI3K-Akt pathway proteins was observed in male neutrophils. Conclusions The study demonstrated the influence of BPA on neutrophil functions associated with locomotion and pathogen elimination, which in turn may disturb the immune response of these cells in both women and men. Analysis of the obtained data showed that the effect of this xenoestrogen on the human neutrophils was more pronounced than E2.

scholarly journals Neuropilin 1 and Neuropilin 2 gene invalidation or pharmacological inhibition reveals their relevance for the treatment of metastatic renal cell carcinoma

2021 ◽  
Vol 40 (1) ◽  
Author(s):  
Aurore Dumond ◽  
Etienne Brachet ◽  
Jérôme Durivault ◽  
Valérie Vial ◽  
Anna K. Puszko ◽  
...  
Keyword(s):  
Renal Cell Carcinoma ◽  
Cell Carcinoma ◽  
Renal Cell ◽  
Cell Metabolism ◽  
Boyden Chamber ◽  
Migration Ability ◽  
Cell Renal Cell Carcinoma ◽  
Immunodeficient Mice ◽  
And Migration

Abstract Background Despite the improvement of relapse-free survival mediated by anti-angiogenic drugs like sunitinib (Sutent®), or by combinations of anti-angiogenic drugs with immunotherapy, metastatic clear cell Renal Cell Carcinoma (mccRCC) remain incurable. Hence, new relevant treatments are urgently needed. The VEGFs coreceptors, Neuropilins 1, 2 (NRP1, 2) are expressed on several tumor cells including ccRCC. We analyzed the role of the VEGFs/NRPs signaling in ccRCC aggressiveness and evaluated the relevance to target this pathway. Methods We correlated the NRP1, 2 levels to patients’ survival using online available data base. Human and mouse ccRCC cells were knocked-out for the NRP1 and NRP2 genes by a CRISPR/Cas9 method. The number of metabolically active cells was evaluated by XTT assays. Migration ability was determined by wound closure experiments and invasion ability by using Boyden chamber coated with collagen. Production of VEGFA and VEGFC was evaluated by ELISA. Experimental ccRCC were generated in immuno-competent/deficient mice. The effects of a competitive inhibitor of NRP1, 2, NRPa-308, was tested in vitro and in vivo with the above-mentioned tests and on experimental ccRCC. NRPa-308 docking was performed on both NRPs. Results Knock-out of the NRP1 and NRP2 genes inhibited cell metabolism and migration and stimulated the expression of VEGFA or VEGFC, respectively. NRPa-308 presented a higher affinity for NRP2 than for NRP1. It decreased cell metabolism and migration/invasion more efficiently than sunitinib and the commercially available NRP inhibitor EG00229. NRPa-308 presented a robust inhibition of experimental ccRCC growth in immunocompetent and immunodeficient mice. Such inhibition was associated with decreased expression of several pro-tumoral factors. Analysis of the TCGA database showed that the NRP2 pathway, more than the NRP1 pathway correlates with tumor aggressiveness only in metastatic patients. Conclusions Our study strongly suggests that inhibiting NRPs is a relevant treatment for mccRCC patients in therapeutic impasses and NRPa-308 represents a relevant hit.

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