Dissociation of murine oral mucosal tissues for single cell applications
J Immunol Methods. 2023 Dec 22:113605. doi: 10.1016/j.jim.2023.113605. Online ahead of print.ABSTRACTSingle-cell RNA sequencing and flow cytometry approaches have been instrumental in understanding cellular states within various tissues and organs. However, tissue dissociation methods can potentially alter results and create bias due to preferential recovery of particular cell types. Here we present efforts to optimize methods for dissociation of murine oral mucosal tissues and provide three different protocols that can be utilized to isolate major cell populations in the oral mucosa. These methods can be used both in heal...
Source: Journal of Immunological Methods - December 24, 2023 Category: Allergy & Immunology Authors: Tomoko Ikeuchi Ramin Akhi Belmaliz Cardona Rodriguez David Fraser Drake Williams Kim Tae Sung Teresa Greenwell-Wild Andrew Overmiller Maria Morasso Niki Moutsopoulos Source Type: research
Novel hapten design, highly sensitive monoclonal antibody production, and immunoassay development for rapid screening of illegally added chloramphenicol in cosmetics
J Immunol Methods. 2023 Dec 22:113604. doi: 10.1016/j.jim.2023.113604. Online ahead of print.ABSTRACTHapten design and synthesis have been regarded as the key factor to generate high-quality antibodies. In the present study, a novel hapten of chloramphenicol was synthesized, characterized and compared with two conventional haptens. The new hapten generated mAb 4B5 showed higher sensitivity and titer than the other two haptens-based mAbs. The haptens synthesized with the structure of chloramphenicol base generated more sensitive antibodies than the hapten with chloramphenicol succinate, and the spacer arm linked to the phen...
Source: Journal of Immunological Methods - December 24, 2023 Category: Allergy & Immunology Authors: Zhaoxiang Wang Mian Wang Xiaoxiang Fu Jingqi Qian Min Wang Guiyu Tan Source Type: research
A SMART method for isolating monoclonal antibodies from individual rhesus macaque memory B cells
J Immunol Methods. 2023 Dec 14;525:113602. doi: 10.1016/j.jim.2023.113602. Online ahead of print.ABSTRACTCharacterizing antigen-specific B cells is a critical component of vaccine and infectious disease studies in rhesus macaques (RMs). However, it is challenging to capture immunoglobulin variable (IgV) genes from individual RM B cells using 5' multiplex (MTPX) primers in nested PCR reactions. In particular, the diversity within RM IgV gene leader sequences necessitates large 5' MTPX primer sets to amplify IgV genes, decreasing PCR efficiency. To address this problem, we developed a switching mechanism at the 5' ends of th...
Source: Journal of Immunological Methods - December 16, 2023 Category: Allergy & Immunology Authors: Jason T Weinfurter Sarah N Bennett Matthew R Reynolds Source Type: research
A SMART method for isolating monoclonal antibodies from individual rhesus macaque memory B cells
J Immunol Methods. 2023 Dec 14;525:113602. doi: 10.1016/j.jim.2023.113602. Online ahead of print.ABSTRACTCharacterizing antigen-specific B cells is a critical component of vaccine and infectious disease studies in rhesus macaques (RMs). However, it is challenging to capture immunoglobulin variable (IgV) genes from individual RM B cells using 5' multiplex (MTPX) primers in nested PCR reactions. In particular, the diversity within RM IgV gene leader sequences necessitates large 5' MTPX primer sets to amplify IgV genes, decreasing PCR efficiency. To address this problem, we developed a switching mechanism at the 5' ends of th...
Source: Journal of Immunological Methods - December 16, 2023 Category: Allergy & Immunology Authors: Jason T Weinfurter Sarah N Bennett Matthew R Reynolds Source Type: research
A SMART method for isolating monoclonal antibodies from individual rhesus macaque memory B cells
J Immunol Methods. 2023 Dec 14;525:113602. doi: 10.1016/j.jim.2023.113602. Online ahead of print.ABSTRACTCharacterizing antigen-specific B cells is a critical component of vaccine and infectious disease studies in rhesus macaques (RMs). However, it is challenging to capture immunoglobulin variable (IgV) genes from individual RM B cells using 5' multiplex (MTPX) primers in nested PCR reactions. In particular, the diversity within RM IgV gene leader sequences necessitates large 5' MTPX primer sets to amplify IgV genes, decreasing PCR efficiency. To address this problem, we developed a switching mechanism at the 5' ends of th...
Source: Journal of Immunological Methods - December 16, 2023 Category: Allergy & Immunology Authors: Jason T Weinfurter Sarah N Bennett Matthew R Reynolds Source Type: research
A SMART method for isolating monoclonal antibodies from individual rhesus macaque memory B cells
J Immunol Methods. 2023 Dec 14:113602. doi: 10.1016/j.jim.2023.113602. Online ahead of print.ABSTRACTCharacterizing antigen-specific B cells is a critical component of vaccine and infectious disease studies in rhesus macaques (RMs). However, it is challenging to capture immunoglobulin variable (IgV) genes from individual RM B cells using 5' multiplex (MTPX) primers in nested PCR reactions. In particular, the diversity within RM IgV gene leader sequences necessitates large 5' MTPX primer sets to amplify IgV genes, decreasing PCR efficiency. To address this problem, we developed a switching mechanism at the 5' ends of the RN...
Source: Journal of Immunological Methods - December 16, 2023 Category: Allergy & Immunology Authors: Jason T Weinfurter Sarah N Bennett Matthew Reynolds Source Type: research
A SMART method for isolating monoclonal antibodies from individual rhesus macaque memory B cells
J Immunol Methods. 2023 Dec 14:113602. doi: 10.1016/j.jim.2023.113602. Online ahead of print.ABSTRACTCharacterizing antigen-specific B cells is a critical component of vaccine and infectious disease studies in rhesus macaques (RMs). However, it is challenging to capture immunoglobulin variable (IgV) genes from individual RM B cells using 5' multiplex (MTPX) primers in nested PCR reactions. In particular, the diversity within RM IgV gene leader sequences necessitates large 5' MTPX primer sets to amplify IgV genes, decreasing PCR efficiency. To address this problem, we developed a switching mechanism at the 5' ends of the RN...
Source: Journal of Immunological Methods - December 16, 2023 Category: Allergy & Immunology Authors: Jason T Weinfurter Sarah N Bennett Matthew Reynolds Source Type: research
A SMART method for isolating monoclonal antibodies from individual rhesus macaque memory B cells
J Immunol Methods. 2023 Dec 14:113602. doi: 10.1016/j.jim.2023.113602. Online ahead of print.ABSTRACTCharacterizing antigen-specific B cells is a critical component of vaccine and infectious disease studies in rhesus macaques (RMs). However, it is challenging to capture immunoglobulin variable (IgV) genes from individual RM B cells using 5' multiplex (MTPX) primers in nested PCR reactions. In particular, the diversity within RM IgV gene leader sequences necessitates large 5' MTPX primer sets to amplify IgV genes, decreasing PCR efficiency. To address this problem, we developed a switching mechanism at the 5' ends of the RN...
Source: Journal of Immunological Methods - December 16, 2023 Category: Allergy & Immunology Authors: Jason T Weinfurter Sarah N Bennett Matthew Reynolds Source Type: research
A SMART method for isolating monoclonal antibodies from individual rhesus macaque memory B cells
J Immunol Methods. 2023 Dec 14:113602. doi: 10.1016/j.jim.2023.113602. Online ahead of print.ABSTRACTCharacterizing antigen-specific B cells is a critical component of vaccine and infectious disease studies in rhesus macaques (RMs). However, it is challenging to capture immunoglobulin variable (IgV) genes from individual RM B cells using 5' multiplex (MTPX) primers in nested PCR reactions. In particular, the diversity within RM IgV gene leader sequences necessitates large 5' MTPX primer sets to amplify IgV genes, decreasing PCR efficiency. To address this problem, we developed a switching mechanism at the 5' ends of the RN...
Source: Journal of Immunological Methods - December 16, 2023 Category: Allergy & Immunology Authors: Jason T Weinfurter Sarah N Bennett Matthew Reynolds Source Type: research
Variegate expression of Cre recombinase in hematopoietic cells in CD11c-cre transgenic mice
This study also suggests that the interpretation of some reports using this CD11c-Cre transgenic mice may need to be re-considered based on a careful evaluation of the cell type-specificity of Cre-mediated in their model.PMID:38101508 | DOI:10.1016/j.jim.2023.113600 (Source: Journal of Immunological Methods)
Source: Journal of Immunological Methods - December 15, 2023 Category: Allergy & Immunology Authors: Claire Murat Sylvie Guerder Source Type: research
Tumor nano-lysate activates dendritic cells to evoke a preventative immune response
This study aimed to gain a better understanding of the immune response behind the success of the vaccine in vivo, through use of ex vivo and in vivo assays. Here, we analyze the activation of various immune cells isolated from healthy mouse spleens and find that antigen-presenting cells (APCs) such as dendritic cells (DCs) are being activated following 24 h incubation with 1:10 mg TNL/mg splenocytes. These cells were further explored to determine the pathway by which activation is occurring, and it was observed that TNL are phagocytosed by DCs to activate NF-kB and c-Fos pathways, resulting in enhanced cytokine release aft...
Source: Journal of Immunological Methods - December 13, 2023 Category: Allergy & Immunology Authors: Jenna A Dombroski Abigail R Fabiano Samantha V Knoblauch Schyler J Rowland Katherine N Gibson-Corley Michael R King Source Type: research
Tumor nano-lysate activates dendritic cells to evoke a preventative immune response
This study aims to gain a better understanding of the immune response behind the success of the vaccine in vivo, through use of ex vivo and in vivo assays. Here, we analyze the activation of various immune cells isolated from healthy mouse spleens and find that antigen-presenting cells (APCs) such as dendritic cells (DCs) are being activated following 24 h incubation with 1:10 mg TNL/mg splenocytes. These cells were further explored to determine the pathway by which activation is occurring, and it was observed that TNL are phagocytosed by DCs to activate NF-kB and c-Fos pathways, resulting in enhanced cytokine release afte...
Source: Journal of Immunological Methods - December 13, 2023 Category: Allergy & Immunology Authors: Jenna A Dombroski Abigail R Fabiano Samantha V Knoblauch Schyler J Rowland Katherine N Gibson-Corley Michael R King Source Type: research
Tumor nano-lysate activates dendritic cells to evoke a preventative immune response
This study aims to gain a better understanding of the immune response behind the success of the vaccine in vivo, through use of ex vivo and in vivo assays. Here, we analyze the activation of various immune cells isolated from healthy mouse spleens and find that antigen-presenting cells (APCs) such as dendritic cells (DCs) are being activated following 24 h incubation with 1:10 mg TNL/mg splenocytes. These cells were further explored to determine the pathway by which activation is occurring, and it was observed that TNL are phagocytosed by DCs to activate NF-kB and c-Fos pathways, resulting in enhanced cytokine release afte...
Source: Journal of Immunological Methods - December 13, 2023 Category: Allergy & Immunology Authors: Jenna A Dombroski Abigail R Fabiano Samantha V Knoblauch Schyler J Rowland Katherine N Gibson-Corley Michael R King Source Type: research
Application of enzyme-linked immunosorbent assay to detect antimicrobial peptides in human intestinal lumen
J Immunol Methods. 2023 Dec 9:113599. doi: 10.1016/j.jim.2023.113599. Online ahead of print.ABSTRACTIntestinal transplantation is the definitive treatment for intestinal failure. However, tissue rejection and graft-versus-host disease are relatively common complications, necessitating aggressive immunosuppression that can itself pose further complications. Tracking intraluminal markers in ileal effluent from standard ileostomies may present a noninvasive and sensitive way to detect developing pathology within the intestinal graft. This would be an improvement compared to current assessments, which are limited by poor sensi...
Source: Journal of Immunological Methods - December 11, 2023 Category: Allergy & Immunology Authors: Julie S Hong Abrar Shamim Hussein Atta Eric B Nonnecke Sarah Merl Satyajit Patwardhan Elin Manell Esad Gunes Philip Jordache Bryan Chen Wuyuan Lu Bo Shen Beatrice Dionigi Ravi P Kiran Megan Sykes Emmanuel Zorn Charles L Bevins Joshua Weiner Source Type: research
Application of enzyme-linked immunosorbent assay to detect antimicrobial peptides in human intestinal lumen
J Immunol Methods. 2023 Dec 9:113599. doi: 10.1016/j.jim.2023.113599. Online ahead of print.ABSTRACTIntestinal transplantation is the definitive treatment for intestinal failure. However, tissue rejection and graft-versus-host disease are relatively common complications, necessitating aggressive immunosuppression that can itself pose further complications. Tracking intraluminal markers in ileal effluent from standard ileostomies may present a noninvasive and sensitive way to detect developing pathology within the intestinal graft. This would be an improvement compared to current assessments, which are limited by poor sensi...
Source: Journal of Immunological Methods - December 11, 2023 Category: Allergy & Immunology Authors: Julie S Hong Abrar Shamim Hussein Atta Eric B Nonnecke Sarah Merl Satyajit Patwardhan Elin Manell Esad Gunes Philip Jordache Bryan Chen Wuyuan Lu Bo Shen Beatrice Dionigi Ravi P Kiran Megan Sykes Emmanuel Zorn Charles L Bevins Joshua Weiner Source Type: research
