scholarly journals A preliminary study to evaluate the immune responses induced by immunization of dogs with inactivated Ehrlichia canis organisms

2005 ◽  
Vol 72 (2) ◽  
Author(s):  
Sunita Mahan ◽  
P.J. Kelly ◽  
S.M. Mahan
Keyword(s):  
Immune Responses ◽  
Western Blotting ◽  
Lymphocyte Proliferation ◽  
Mononuclear Cells ◽  
Antibody Responses ◽  
Ehrlichia Canis ◽  
Intracellular Pathogen ◽  
Peripheral Blood Mononuclear ◽  
Canine Monocytic Ehrlichiosis ◽  
Antibody Titres

Ehrlichia canis is an intracellular pathogen that causes canine monocytic ehrlichiosis. Although the role of antibody responses cannot be discounted, control of this intracellular pathogen is expected to be by cell mediated immune responses. The immune responses in dogs immunized with inactivated E. canis organisms in combination with Quil A were evaluated. Immunization provoked strong humoral and cellular immune responses, which were demonstrable by Western blotting and lymphocyte proliferation assays. By Western blotting antibodies to several immunodominant E. canis proteins were detected in serum from immunized dogs and antibody titres increased after each immunization. The complement of immunogenic proteins recognized by the antisera were similar to those recognized in serum from infected dogs. Upon challenge with live E. canis, rapid anamnestic humoral responses were detected in the serum of immunized dogs and primary antibody responses were detected in the serum from control dogs. Following immunization, a lymphocyte proliferative response (cellular immunity) was detected in peripheral blood mononuclear cells (PBMNs) of immunized dogs upon stimulation with E. canis antigens. These responses were absent from non-immunized control dogs until after infection with live E. canis, when antigen specific-lymphocyte proliferation responses were also detected in the PBMNs of the control dogs. It can be thus concluded that immunization against canine monocytic ehrlichiosis may be feasible. However, the immunization regimen needs to be optimized and a detailed investigation needs to be done to determine if this regimen can prevent development of acute and chronic disease.

Traditional Acupuncture Increases the Content of Beta-Endorphin in Immune Cells and Influences Mitogen Induced Proliferation

1991 ◽  
Vol 19 (02) ◽  
pp. 101-104 ◽  
Author(s):  
Mauro Bianchi ◽  
Edda Jotti ◽  
Paola Sacerdote ◽  
Alberto E. Panerai
Keyword(s):  
Immune Responses ◽  
T Lymphocyte ◽  
Peripheral Blood Mononuclear Cells ◽  
Immune Cells ◽  
Lymphocyte Proliferation ◽  
Mononuclear Cells ◽  
Peripheral Blood Mononuclear ◽  
Beta Endorphin ◽  
T Lymphocyte Proliferation ◽  
Blood Mononuclear Cells

We measured beta-endorphin concentrations in peripheral blood mononuclear cells and mitogen-induced T-lymphocyte proliferation in patient who underwent treatment with traditional acupuncture. Traditional acupuncture increased both the concentrations of the opioid in the immune cells and lymphocyte proliferation. Our data are consistent with the hypothesis that traditional acupuncture modulates immune responses in man.

Sipuleucel-T (sip-T)–induced proliferative CD8+ T-cell responses to immunizing and secondary antigens.

2016 ◽  
Vol 34 (2_suppl) ◽  
pp. 165-165 ◽  
Author(s):  
Emmanuel S. Antonarakis ◽  
David I. Quinn ◽  
Adam S. Kibel ◽  
Daniel Peter Petrylak ◽  
Nancy N. Chang ◽  
...  
Keyword(s):  
T Cell ◽  
Immune Responses ◽  
Mononuclear Cells ◽  
T Cell Responses ◽  
Cd8 T Cell ◽  
Antibody Responses ◽  
Castration Resistant Prostate Cancer ◽  
Peripheral Blood Mononuclear ◽  
Cell Responses

165 Background: Sip-T is an FDA-approved autologous immunotherapy for patients (pts) with asymptomatic or minimally symptomatic metastatic castration-resistant prostate cancer (mCRPC). Sip-T is manufactured from peripheral blood mononuclear cells cultured with PA2024, a fusion antigen of prostatic acid phosphatase (PAP) conjugated to granulocyte macrophage-colony stimulating factor (GM-CSF). In sip-T–treated pts, T cell and antibody responses to PA2024 or PAP as well as antibody responses to secondary antigens (i.e., antigen spread) correlate with improved overall survival. To explore the biology of this relationship, we further characterized the T cell subpopulations involved in the cellular immune responses to sip-T. Methods: In vitro proliferative CD8+ (cytotoxic T lymphocyte) and CD4+ (T helper) T cell responses to PA2024 and PAP as well as to secondary antigens (PSA, LGALS3, and KRAS) were evaluated using flow cytometry on pt samples from two sip-T–containing clinical trials (STAND [NCT01431391] and STRIDE [NCT01981122]). Results: PA2024-specific CD8+ and CD4+ responses were observed beginning 2 weeks after the first sip-T infusion through week 26 in most evaluable pts. CD8+ and CD4+ responses to PAP were also observed, although the magnitude of this response (to a self-antigen) was smaller when compared with PA2024 responses. Most pts with CD8+ responses to PA2024 also had a CD4+ response, which occurred more frequently than CD8+ responses. Both CD8+ and CD4+ responses to secondary antigens were amplified after sip-T, and these CD8+ proliferative responses to secondary antigens were greater in magnitude compared with CD4+ responses. Conclusions: Here,we report the first evidence of antigen-specific CD8+ responses in pts receiving sip-T, indicating CD8+ T cell involvement in sip-T–mediated immune responses that occur in concert with the expected CD4+ T cell responses to soluble antigens (i.e., PA2024). These CD8+ responses were durable, lasting up to 26 weeks post–sip-T treatment. Importantly, responses to secondary antigens and in particular, CD8+ responses, were also amplified after sip-T treatment, suggesting that antigen spread could be resulting from sip-T–mediated tumor cell lysis. Clinical trial information: NCT01431391 and NCT01981122.

scholarly journals Viability and Functional Activity of Cryopreserved Mononuclear Cells

2000 ◽  
Vol 7 (4) ◽  
pp. 714-716 ◽  
Author(s):  
Adriana Weinberg ◽  
Li Zhang ◽  
Darby Brown ◽  
Alejo Erice ◽  
Bruce Polsky ◽  
...  
Keyword(s):  
Clinical Trial ◽  
Functional Activity ◽  
Lymphocyte Proliferation ◽  
Mononuclear Cells ◽  
Cell Number ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells ◽  
And Function ◽  
Functional Analyses ◽  
Cd4 Cell

ABSTRACT Factors that influence viability and function of cryopreserved peripheral blood mononuclear cells (PBMC) were identified on 54 samples from 27 AIDS Clinical Trial Units. PBMC viability ranged from 1 to 96% with a median of 70%, was higher in laboratories with experienced staff, and was not significantly associated with CD4 cell number. Function of cryopreserved PBMC, measured by lymphocyte proliferation, was associated with viability. Preparations with viability greater than or equal to 70% had consistent proliferative responses and were suitable for functional analyses.

scholarly journals P362 Longitudinal antibody response to SARS-CoV-2 infection and antibody responses to COVID-19 vaccination in Inflammatory Bowel Disease patients receiving biologics

2021 ◽  
Vol 15 (Supplement_1) ◽  
pp. S380-S381
Author(s):  
S Y Wong ◽  
R Dixon ◽  
S Gold ◽  
M Vicky ◽  
D Helmus ◽  
...  
Keyword(s):  
New York ◽  
Immune Responses ◽  
Antibody Responses ◽  
Large Hospital ◽  
Nucleocapsid Proteins ◽  
Medication Effects ◽  
Vaccine Trials ◽  
Antibody Titres ◽  
Inflammatory Bowel ◽  
Hospital Center

Abstract Background Given that IBD patients were excluded from COVID-19 vaccine trials, there is a lack of vaccine efficacy data in this population. In this study, we evaluated longitudinal serological responses to SARS-CoV-2 infection as well as to COVID-19 vaccination in IBD patients. Methods We collected clinical data and sera from IBD patients enrolled in an observational SARS-CoV-2 sero-surveillance study at our large hospital center in New York City during routine infusions and clinic visits. To distinguish between infection and vaccination, sera was collected prior to vaccination where possible, and all sera was tested for both antibodies to SARS-CoV-2-specific RBD, the target of current available vaccines in the U.S., and nucleocapsid proteins. Results Our results reveal waning antibody titres in 13 of 16 (81%) patients infected with SARS-CoV-2 over a course of 6-7 months. Of 48 vaccinated patients, 16 patients completed vaccine schedules with two doses, and all 16 (100%) achieved seroconversion above the threshold required for convalescent plasma donation. Conclusion While antibody responses to infection in IBD patients have questionable stability, completion of the COVID-19 vaccine series in IBD patients results in robust serological responses. To our knowledge is the first data confirming adequate serological responses to COVID-19 vaccination in IBD patients with and without biologic medications. Studies are needed to assess adequacy of dosing schedules, medication effects, measurement of cell-mediated responses, durability of immune responses, and clinical efficacy of COVID-19 vaccines in IBD patients.

scholarly journals Cytokine Profiles and Cell Proliferation Responses to Truncated ORF2 Protein in Iranian Patients Recovered from Hepatitis E Infection

2015 ◽  
Vol 2015 ◽  
pp. 1-7 ◽  
Author(s):  
Reza Taherkhani ◽  
Fatemeh Farshadpour ◽  
Manoochehr Makvandi ◽  
Hamid Rajabi Memari ◽  
Ali Reza Samarbafzadeh ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Immune Responses ◽  
Mononuclear Cells ◽  
Hepatitis E ◽  
Control Group ◽  
Peripheral Blood Mononuclear ◽  
Orf2 Protein ◽  
Cellular Immune ◽  
Iranian Patients

Background.The aim of this study was to evaluatehepatitis E virus(HEV) specific cellular immune responses to truncated ORF2 protein in Iranian patients recovered from HEV infection. Information about HEV-specific immune responses could be useful in finding an effective way for development of HEV vaccine.Methods.A truncated form of HEV ORF2 protein containing amino acids 112-608 was used to stimulate peripheral blood mononuclear cells (PBMCs) separated from HEV-recovered and control groups. Finally, the levels of four cytokines, IFN-γELISPOT, and cell proliferative responses following stimulation with the truncated ORF2 protein were assessed in the both groups.Results.The truncated ORF2 protein was able to induce IFN-γELISPOT and cell proliferation responses and to produce significant amounts of IFN-γand IL-12 cytokines, but low amounts of IL-10 and IL-4 cytokinesin vitro. These responses were significantly higher in the recovered group compared to the control group. These results indicate the antigenic nature of the truncated ORF2 protein and production of T helper type 1 cytokines.Conclusion.The truncated ORF2 protein can effectively induce significant cellular immune responsesand can be introduced as a potential vaccine candidate. However, further studies are required to evaluate this proteinin vivo.

Clinical significance of insulin peptide-specific interferon-γ-related immune responses in ketosis-prone type 2 diabetes

2021 ◽  
Author(s):  
Atsushi Satomura ◽  
Yoichi Oikawa ◽  
Akifumi Haisa ◽  
Seiya Suzuki ◽  
Shunpei Nakanishi ◽  
...  
Keyword(s):  
Type 2 Diabetes ◽  
Immune Responses ◽  
Elispot Assay ◽  
Mononuclear Cells ◽  
Cell Function ◽  
Β Cell ◽  
Peripheral Blood Mononuclear ◽  
Β Cell Function ◽  
Ifn Γ

Abstract Context Unprovoked A−β+ ketosis-prone type 2 diabetes (KPD) is characterized by the sudden onset of diabetic ketosis/ketoacidosis (DK/DKA) without precipitating factors, negative anti-islet autoantibodies (“A−”), and preservation of β-cell function (“β+”) after recovery from DKA. Although this phenotype often appears with acute hyperglycemia and DK/DKA just like acute-onset type 1 diabetes (AT1D), the involvement of anti-islet immune responses remains unknown. Objective We sought to clarify the immunological role of insulin-associated molecules in unprovoked A−β+ KPD. Methods In this cross-sectional study, blood samples from 75 participants (42 with AT1D and 33 with KPD) were evaluated for interferon (IFN)-γ-secreting peripheral blood mononuclear cells (PBMCs) reactive to four insulin B-chain amino acid 9–23-related peptides (B:9–23rPep) using an enzyme-linked immunospot (ELISpot) assay. Results Overall, 36.4% (12/33) of KPD participants showed positive IFN-γ ELISpot assay results; the positivity rate in KPD was similar to that in AT1D (38.1%; 16/42) and significantly higher than the previously reported rate in type 2 diabetes (8%; 2/25; P < 0.0167). Moreover, B:9–23rPep-specific IFN-γ-producing PBMC frequency was negatively correlated with age and ad lib serum C-peptide levels in all KPD participants and positively correlated with HbA1c level in KPD participants with positive IFN-γ ELISpot results. Conclusions These findings suggest the involvement of B:9–23rPep-specific IFN-γ-related immunoreactivity in the pathophysiology of some unprovoked A−β+ KPD. Moreover, increased immunoreactivity may reflect transiently decreased β-cell function and increased disease activity at the onset of DK/DKA, thereby playing a key role in DK/DKA development in this KPD phenotype.

scholarly journals Mapping glycan-mediated galectin-3 interactions by live cell proximity labeling

2020 ◽  
Vol 117 (44) ◽  
pp. 27329-27338
Author(s):  
Eugene Joeh ◽  
Timothy O’Leary ◽  
Weichao Li ◽  
Richard Hawkins ◽  
Jonathan R. Hung ◽  
...  
Keyword(s):  
Immune Responses ◽  
Hepatic Stellate Cells ◽  
Mononuclear Cells ◽  
Three Dimensional ◽  
Stellate Cells ◽  
Live Cells ◽  
Quantitative Mass Spectrometry ◽  
Peripheral Blood Mononuclear ◽  
Galectin 3 ◽  
Labeling Method

Galectin-3 is a glycan-binding protein (GBP) that binds β-galactoside glycan structures to orchestrate a variety of important biological events, including the activation of hepatic stellate cells and regulation of immune responses. While the requisite glycan epitopes needed to bind galectin-3 have long been elucidated, the cellular glycoproteins that bear these glycan signatures remain unknown. Given the importance of the three-dimensional (3D) arrangement of glycans in dictating GBP interactions, strategies that allow the identification of GBP receptors in live cells, where the native glycan presentation and glycoprotein expression are preserved, have significant advantages over static and artificial systems. Here we describe the integration of a proximity labeling method and quantitative mass spectrometry to map the glycan and glycoprotein interactors for galectin-3 in live human hepatic stellate cells and peripheral blood mononuclear cells. Understanding the identity of the glycoproteins and defining the structures of the glycans will empower efforts to design and develop selective therapeutics to mitigate galectin-3–mediated biological events.

scholarly journals Mesenchymal stem/stromal cell quality control: validation of mixed lymphocyte reaction assay using flow cytometry according to ICH Q2(R1)

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Tess Nicotra ◽  
Aurélie Desnos ◽  
Justine Halimi ◽  
Hélène Antonot ◽  
Loïc Reppel ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Lymphocyte Proliferation ◽  
Mononuclear Cells ◽  
Line Graph ◽  
Area Under The Curve ◽  
Measurement Range ◽  
Immunomodulatory Activity ◽  
Peripheral Blood Mononuclear ◽  
Wide Range ◽  
Inhibition Control

Abstract Background Mesenchymal stem/stromal cells (MSC) have immunomodulatory properties, studied in a wide range of diseases. Validated quality controls must confirm this activity in the context of clinical trials. This study presents a method’s validation, assessing MSC’s ability to inhibit lymphocyte proliferation, according to the ICH Q2 standard. Methods MSC were co-cultured with CellTrace™ Violet-labeled Peripheral blood mononuclear cells (PBMC) coming from a bank of ten donors, at seven different ratios for 7 days. Cell trace violet PBMC bank was validated in parallel. Flow cytometry analysis was used to obtain the division percentage of T cells. The percentage of inhibition of lymphocyte proliferation by MSC, for each ratio X, was calculated using the formula: Ratio × percentage of inhibition = (control percentage of division—ratio × percentage of division)/control percentage of division. The inhibition percentage of lymphocyte proliferation function of co-culture ratios was represented in a line graph. The corresponding area under the curve was calculated, representing MSC’s ability to inhibit lymphocyte proliferation. Results Two cell trace violet PBMC banks were compared for bank validation. When compared using four different MSC samples coming each from a different donor, their area under the curve did not show any statistical differences and were correlated. Moreover, the stability of one cell trace violet PBMC bank was confirmed up to 509 days of storage. Analytical parameters were investigated for method validation. Analysis of repeatability and reproducibility respectively showed a standard deviation of 6.1% and 4.6%. The assay was robust regarding PBMC, as no statistical differences were found between inhibitory activities when testing three adjacent concentrations of PBMC. Still, attention is needed on MSC quantity as it can influence results. Linearity was evaluated: the percentage of inhibition of lymphocyte proliferation function of co-culture ratios was linear on the exploited range. Finally, the assay measurement range allowed to differentiate MSC presenting different inhibition activities. Conclusion This quantification method displayed low analytical variability and no inter-bank variability of PBMC. However, MSC quantification should be checked before co-culture to reduce variability. Therefore, it could be used for the qualification of MSC batches’ immunomodulatory activity.

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