s mitis uf28 nd  (ATCC)

Journal: Microbiology Spectrum

Article Title: Deficient Complement Opsonization Impairs Mycobacterium avium Killing by Neutrophils in Cystic Fibrosis

doi: 10.1128/spectrum.03279-22

Figure Lengend Snippet: Complement and antibody depletion and neutrophil killing of M. avium . (A) Healthy donor neutrophils were incubated with M. avium opsonized with healthy donor heat-inactivated serum (HS), intact healthy donor serum (WS), or WS depleted (dep) of complement C3, C1q, or factor B (FB) for 60 min. Surviving M. avium colonies were compared to the corresponding t = 0 inoculum to calculate percentage of killing ( n = 10 to 12). ****, P ≤ 0.0001 by one-way analysis of variance (ANOVA). (B) Healthy donor neutrophils were incubated with M. avium opsonized with healthy donor HP, HP treated with protein A- or protein G-agarose, and HP depleted of IgM for 60 min. Surviving M. avium colonies were compared to the corresponding t = 0 inoculum to calculate percentage of killing ( n = 7 to 9). (C) Healthy donor neutrophils were incubated with M. avium opsonized with WP, WP treated with protein A- or protein G-agarose, and WP depleted of IgM for 60 min. Surviving M. avium colonies were compared to the corresponding t = 0 inoculum to calculate percentage of killing ( n = 7 to 9). ***, P = 0.0003; **, P = 0.019 by one-way ANOVA.

Article Snippet: The blots were probed with anti-C3 (BioLegend, 846302), anti-C1q (Novus NB100-64420), anti-IgA (Southern Biotech 2050-05), anti-IgG (Abcam ab97175), or anti-IgM (Southern Biotech 2020-05), as indicated, overnight.

Techniques: Incubation

Journal: Microbiology Spectrum

Article Title: Deficient Complement Opsonization Impairs Mycobacterium avium Killing by Neutrophils in Cystic Fibrosis

doi: 10.1128/spectrum.03279-22

Figure Lengend Snippet: Complement and antibody opsonization of M. avium . Comparison of protein levels of C3, C1q, IgG, IgA, and IgM bound to M. avium (Mav) opsonized with WP compared to WP alone via Western blot. C3 and IgM (M) were both found to associate with M. avium opsonized with WP. C1q, IgG (G), and IgA (A) were also associated with opsonized M. avium in but in small amounts.

Article Snippet: The blots were probed with anti-C3 (BioLegend, 846302), anti-C1q (Novus NB100-64420), anti-IgA (Southern Biotech 2050-05), anti-IgG (Abcam ab97175), or anti-IgM (Southern Biotech 2020-05), as indicated, overnight.

Techniques: Comparison, Western Blot

Journal: Microbiology Spectrum

Article Title: Deficient Complement Opsonization Impairs Mycobacterium avium Killing by Neutrophils in Cystic Fibrosis

doi: 10.1128/spectrum.03279-22

Figure Lengend Snippet: Complement levels in persons with CF (pwCF) with and without nontuberculous mycobacteria (NTM) infection. (A) Plasma levels of complement factors C1q, C3, C3b/iC3b, C4, factor B, and factor H in healthy donors, pwCF with no history of NTM infection (CF −NTM), and pwCF with a history of NTM infection (CF +NTM). Complement C3 and C3b/iC3b levels were significantly higher in pwCF with a history of NTM compared to pwCF without a history of NTM ( P = 0.01 and P = 0.02, respectively, by one-way ANOVA) and were higher than in healthy donors. Complement C4 levels were also higher in pwCF with a history of NTM compared to those without ( P = 0.045), but levels were similar to those of healthy donors. (B) Hierarchical clustering of individual subject complement profiles. Unsupervised clustering of complement profiles revealed two primary clusters representing low (cluster A) and high (cluster B) levels. All HD subjects and all CF without NTM fell within cluster A, along with 5 of 16 (31%) of CF +NTM subjects. Cluster B was comprised entirely of CF +NTM subjects ( P = 0.0003 by two-tail Fisher’s exact test, for subjects with CF).

Article Snippet: The blots were probed with anti-C3 (BioLegend, 846302), anti-C1q (Novus NB100-64420), anti-IgA (Southern Biotech 2050-05), anti-IgG (Abcam ab97175), or anti-IgM (Southern Biotech 2020-05), as indicated, overnight.

Techniques: Infection

Journal: International Journal of Molecular Sciences

Article Title: Heat-Inactivation of Human Serum Destroys C1 Inhibitor, Pro-motes Immune Complex Formation, and Improves Human T Cell Function

doi: 10.3390/ijms22052646

Figure Lengend Snippet: Effects of serum heat-inactivation on complement-bearing immune complexes, complement (co-)factors, and regulators. ( Center ) The depicted scheme represents the initiating steps and essential (co-)factors and regulators of the classical and alternative complement cascade as well as the cleavage pathway of complement factor C3. ( A – G ) Concentrations of complement-bearing immune complexes ( A ) C1q-IgG and ( B ) C3d-IgG. ( C ) The concentration and ( D ) activity of complement regulator C1 inhibitor as well as the concentration of complement (co-)factors ( E ) C3c, ( F ) C4, and ( G ) factor B were determined in native sera (blue dots) and after heat-inactivation (red dots). (median of (A) native n = 17, heat-inactivated (HI) n = 16; (B,E,F,G) n =11; (C) native n = 15, HI n = 14; (D) native n = 8, HI n = 7; Wilcoxon test; * p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: In order to induce complex formation with serum contained C1q and therewith imitate the effect of heat-inactivation, we added a final concentration of 50 µg/mL anti-human anti-C1q antibody (NB100-64420, Novus Biologicals, Centennial, CO, USA) to cells cultured in native sera.

Techniques: Concentration Assay, Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: Heat-Inactivation of Human Serum Destroys C1 Inhibitor, Pro-motes Immune Complex Formation, and Improves Human T Cell Function

doi: 10.3390/ijms22052646

Figure Lengend Snippet: Effects of heat-inactivation are not related to Fcγ receptors II (CD32) and III (CD16) expression but are partially reverted by C1 inhibitor and mimicked by anti-C1q antibody supplementation. ( A , B ) Histogram plots display expression of CD16, CD32, and CD64 (opaque colors) compared to isotype controls (black) and unstained cells (transparent color) in native (A) and HI cultured cells (B) after 48 h of stimulation. One representative experiment is shown. ( C , D ) ΔMFI (difference of MFI antibody and MFI isotype) of CD16 ( C ) and CD32 ( D ) is depicted in a time-dependent manner. Shown is the median ± interquartile range of n = 4 (24 h/48 h/72 h/6 d) and n = 2 (0 h). ( E – H ) C1 inhibitor was added to T cells cultured with heat inactivated serum. In the presence of C1 inhibitor IFNγ ( E ), TNF ( F ), CD28 ( G ), and CD69 ( H ) were analyzed at indicated time points. (Shown is the median of n = 6, each dot represents one donor, Friedman test, post-hoc Dunn’s, * p < 0.05, ** p < 0.01). ( I – K ) Anti-C1q antibody was added to T cells cultured with native sera. TNF secretion ( I ) was measured by ELISA and CD28 ( J ) and CD69 ( K ) expression was analyzed by flow cytometry at indicated time points. (Shown is the median of n = 13; Friedman test, post-hoc Dunn’s, * p < 0.05 ** p < 0.01; *** p < 0.001).

Article Snippet: In order to induce complex formation with serum contained C1q and therewith imitate the effect of heat-inactivation, we added a final concentration of 50 µg/mL anti-human anti-C1q antibody (NB100-64420, Novus Biologicals, Centennial, CO, USA) to cells cultured in native sera.

Techniques: Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Heat-Inactivation of Human Serum Destroys C1 Inhibitor, Pro-motes Immune Complex Formation, and Improves Human T Cell Function

doi: 10.3390/ijms22052646

Figure Lengend Snippet: Schematic illustration of the effects of HI, HI + C1inh, and native + anti-C1q on human CD4+ T cells compared to cells in native serum. Effects are analyzed after 72 h (HI, HI + C1inh) and 48 h (native + anti-C1q), respectively. Each arrow represents a significantly increased (↑), significantly decreased (↓) effect or no significant effect (↔). Effects being also induced by the carrier solution are depicted as no significant effect.

Article Snippet: In order to induce complex formation with serum contained C1q and therewith imitate the effect of heat-inactivation, we added a final concentration of 50 µg/mL anti-human anti-C1q antibody (NB100-64420, Novus Biologicals, Centennial, CO, USA) to cells cultured in native sera.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Heat-Inactivation of Human Serum Destroys C1 Inhibitor, Pro-motes Immune Complex Formation, and Improves Human T Cell Function

doi: 10.3390/ijms22052646

Figure Lengend Snippet: Tests, equipment, and methods used for determination of heat-induced changes.

Article Snippet: In order to induce complex formation with serum contained C1q and therewith imitate the effect of heat-inactivation, we added a final concentration of 50 µg/mL anti-human anti-C1q antibody (NB100-64420, Novus Biologicals, Centennial, CO, USA) to cells cultured in native sera.

Techniques: Activity Assay, Binding Assay, Concentration Assay, Electrophoresis

Journal: Annals of Work Exposures and Health

Article Title: Evaluation of AccuFIT 9000: A Novel Apparatus for Quantitative Fit Testing of Particulate Respirators

doi: 10.1093/annweh/wxaa116

Figure Lengend Snippet: Respirators used in the study.

Article Snippet: , JACKSON SAFETY 64420 P95, SureWerx USA Inc., Elgin, IL, USA a , One size.

Techniques: