scholarly journals A commercial ELISA for detection of interferon gamma in white rhinoceros

2019 ◽  
Vol 31 (4) ◽  
pp. 531-536 ◽  
Author(s):  
Josephine Chileshe ◽  
Wynand J. Goosen ◽  
Peter E. Buss ◽  
Paul D. van Helden ◽  
Robin Warren ◽  
...  
Keyword(s):  
Interferon Gamma ◽  
Incubation Temperature ◽  
Disease Process ◽  
Optimal Recovery ◽  
Enzyme Linked Immunosorbent Assay ◽  
Limit Of Quantification ◽  
White Rhinoceros ◽  
Elisa Plate ◽  
Ifn Γ ◽  
And Control

Bovine tuberculosis (bTB), caused by Mycobacterium bovis, is endemic in Kruger National Park, South Africa, home to the largest population of white rhinoceros ( Ceratotherium simum) in the world. In 2016, the first cases of naturally occurring bTB were reported in white rhinoceros; however, there is a lack of understanding of infection and disease process in this species. Prevention and control of transmission depends on the availability of accurate tools to detect M. bovis infection. Interferon gamma (IFN-γ) assays are a reliable detection method for TB in other animal species, and studies have indicated that these tests can be used in white rhinoceros. We sought to screen and optimize a commercial IFN-γ enzyme-linked immunosorbent assay (ELISA) to detect endogenous white rhinoceros IFN-γ in mitogen-stimulated whole blood as a basis for developing a test for M. bovis infection. Optimizations included identifying ELISA antibodies and determining the effect of sample matrix, ELISA plate incubation temperature, ELISA linearity, assay reproducibility, and the assay’s limit of quantification. The optimized assay employed an equine IFN-γ antibody pair that was used to create a commercial ELISA kit. This ELISA had a linear response to recombinant equine and endogenous rhinoceros IFN-γ (range: 7.8–125 pg/mL). When incubated at 37°C, the ELISA was highly reproducible, with an optimal recovery and a low limit of quantification, indicating that the Mabtech equine IFN-γ ELISAPRO kit is a robust assay for measuring white rhinoceros IFN-γ.

scholarly journals Immediate Interferon Gamma Induction Determines Murine Host Compatibility Differences between Toxoplasma gondii and Neospora caninum

2020 ◽  
Vol 88 (4) ◽  
Author(s):  
Rachel S. Coombs ◽  
Matthew L. Blank ◽  
Elizabeth D. English ◽  
Yaw Adomako-Ankomah ◽  
Ifeanyi-Chukwu Samuel Urama ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Immune Responses ◽  
Interferon Gamma ◽  
Neospora Caninum ◽  
Ectopic Expression ◽  
Parasite Burden ◽  
Interleukin 12 ◽  
Content Type ◽  
Ifn Γ ◽  
And Control

ABSTRACT Rodents are critical for the transmission of Toxoplasma gondii to the definitive feline host via predation, and this relationship has been extensively studied as a model for immune responses to parasites. Neospora caninum is a closely related coccidian parasite of ruminants and canines but is not naturally transmitted by rodents. We compared mouse innate immune responses to N. caninum and T. gondii and found marked differences in cytokine levels and parasite growth kinetics during the first 24 h postinfection (hpi). N. caninum-infected mice produced significantly higher levels of interleukin-12 (IL-12) and interferon gamma (IFN-γ) by as early as 4 hpi, but the level of IFN-γ was significantly lower or undetectable in T. gondii-infected mice during the first 24 hpi. “Immediate” IFN-γ and IL-12p40 production was not detected in MyD88−/− mice. However, unlike IL-12p40−/− and IFN-γ−/− mice, MyD88−/− mice survived N. caninum infections at the dose used in this study. Serial measures of parasite burden showed that MyD88−/− mice were more susceptible to N. caninum infections than wild-type (WT) mice, and control of parasite burdens correlated with a pulse of serum IFN-γ at 3 to 4 days postinfection in the absence of detectable IL-12. Immediate IFN-γ was partially dependent on the T. gondii mouse profilin receptor Toll-like receptor 11 (TLR11), but the ectopic expression of N. caninum profilin in T. gondii had no impact on early IFN-γ production or parasite proliferation. Our data indicate that T. gondii is capable of evading host detection during the first hours after infection, while N. caninum is not, and this is likely due to the early MyD88-dependent recognition of ligands other than profilin.

scholarly journals Changes in interleukin-27 levels in patients with acute coronary syndrome and their clinical significance

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e5652 ◽  
Author(s):  
Lin Zhang ◽  
Junfeng Zhang ◽  
Shaohong Su ◽  
Suyan Luo
Keyword(s):  
Acute Coronary Syndrome ◽  
Angina Pectoris ◽  
Th17 Cells ◽  
Serum Levels ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Groups ◽  
Coronary Syndrome ◽  
Significant Difference ◽  
Ifn Γ ◽  
And Control

Background This study evaluated changes in interleukin (IL)-27 levels in patients with acute coronary syndrome (ACS) and their influence on Th1, Th2, and Th17 cells. Methods Serum levels of IL-27, IL-4, IL-17, and interferon (IFN)-γ in healthy subjects as well as patients with ACS, including stable angina pectoris (SA), unstable angina pectoris (UA), and acute myocardial infarction (AMI), were determined using an enzyme-linked immunosorbent assay. The proportions of Th1, Th2, and Th17 cells among peripheral blood mononuclear cells (PBMCs), were measured using flow cytometry, after incubation with phorbol myristate acetate (PMA) for 4 h. The proportions of Th1 and Th17 cells among PBMCs in AMI and UA were detected after stimulation with IL-27 or PMA + IL-27 for 4, 8, and 12 h. Results Serum levels of IL-27 in patients with AMI and UA were significantly lower than those in SA and control groups, while serum levels of IL-17 and IFN-γ in AMI and UA groups were dramatically increased compared to those in SA and healthy control groups. However, there were no statistically significant differences in serum IL-4. The proportions of Th1 and Th17 cells among PBMCs were statistically significantly higher in the AMI and UA groups than those in the SA and control groups, while there was no statistically significant difference in the proportion of Th2 cells among different groups. For patients with AMI and UA, the effect of co-stimulation of PBMCs with PMA and IL-27 was not significantly different from that of PMA single stimulation, while PMA + IL-27 co-stimulation lowered the Th17 cell proportion significantly compared to PMA single stimulation. Discussion Compared to SA patients and healthy controls, patients with ACS (AMI + UA) had lower serum levels of IL-27 and higher proportions of PBMC Th1 and Th17 cells, which could be attributed to the inhibitory effects of IL-27 on the proliferation of Th17 cells. These results indicated that IL-27 could be a novel therapeutic target in ACS patients.

scholarly journals Latticed Gold Nanoparticle Conjugation via Monomeric Streptavidin in Lateral Flow Assay for Detection of Autoantibody to Interferon-Gamma

Diagnostics ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 987
Author(s):  
Weeraya Thongkum ◽  
Umpa Yasamut ◽  
Koollawat Chupradit ◽  
Supachai Sakkhachornphop ◽  
Jiraprapa Wipasa ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Interferon Gamma ◽  
Colloidal Gold ◽  
Enzyme Linked Immunosorbent Assay ◽  
Adult Onset ◽  
Elisa Titer ◽  
Mycobacterial Diseases ◽  
Ifn Γ ◽  
Immunodeficiency Syndrome ◽  
Specific Symptoms

Adult-onset immunodeficiency syndrome (AOID) patients with autoantibodies (autoAbs) against interferon-gamma (IFN-γ) generally suffer from recurrent and recalcitrant disseminated non-tuberculous mycobacterial diseases. Since the early stages of AOID do not present specific symptoms, diagnosis and treatment of the condition are not practical. A simplified diagnostic method for differentiating AOID from other immunodeficiencies, such as HIV infection, was created. Anti-IFN-γ is generally identified using enzyme-linked immunosorbent assay (ELISA), which involves an instrument and a cumbersome process. Recombinant IFN-γ indirectly conjugated to colloidal gold was used in the modified immunochromatographic (IC) strips. The biotinylated-IFN-γ was incorporated with colloidal-gold-labeled 6HIS-maltose binding protein-monomeric streptavidin (6HISMBP-mSA) and absorbed at the conjugate pad. The efficacy of the IC strip upon applying an anti-IFN-γ autoAb cut-off ELISA titer of 2500, the sensitivity and specificity were 84% and 90.24%, respectively. When a cut-off ELISA titer of 500 was applied, the sensitivity and specificity were 73.52% and 100%, respectively.

scholarly journals The secret behind peripheral inflammation and depression: The hypothalamus

2020 ◽  
Author(s):  
Bruno Pedraz Petrozzi ◽  
Carlo Blecker ◽  
Elena Neumann ◽  
Gebhard Sammer
Keyword(s):  
Mixed Models ◽  
Enzyme Linked Immunosorbent Assay ◽  
Resonance Spectroscopy ◽  
Peripheral Inflammation ◽  
Tnf Α ◽  
Ifn Γ ◽  
And Control ◽  
Hpa Axis Activity ◽  
Inflammatory Component

Abstract In recent years there has been increasing evidence of an inflammatory component due to overstimulation of the hypothalamic-pituitary-adrenals (HPA) in depression. The glutamate metabolites (glutamate and glutamine) are important metabolites that are involved in this stimulation. The aim of this study was to determine the concentrations of hypothalamic glutamate metabolites in depression and to investigate their relationship to peripheral inflammation. Participants with diagnosed depression (DE; n = 24) and control subjects without depression (HC; n = 25) were investigated. Hypothalamic glutamate metabolites were recorded using in vivo magnetic resonance spectroscopy. Peripheral cytokines (IL-6, TNF-α, IFN-γ and IL-1β) were assessed using Enzyme-Linked Immunosorbent Assay. For statistical analysis, generalized mixed models were computed using Poisson distributions and a log link function. The results show overall higher hypothalamic glutamate metabolites in DE compared to HC. High TNF-a and IL-1ß concentrations are associated with high hypothalamic glutamate metabolites in DE. These results provide initial evidence that, in depression, increased HPA axis activity is associated with peripheral inflammation favored by hypothalamic glutamate metabolites.

scholarly journals Predictors of the Therapeutic Response to Intralesional Bivalent HPV Vaccine in Wart Immunotherapy

Vaccines ◽  
2021 ◽  
Vol 9 (11) ◽  
pp. 1280
Author(s):  
Noha M. Hammad ◽  
Ayman Marei ◽  
Gamal El-Didamony ◽  
Zeinb Mortada ◽  
Mona Elradi ◽  
...  
Keyword(s):  
Hpv Vaccine ◽  
Ex Vivo ◽  
Hpv Infection ◽  
Interleukin 4 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Anogenital Warts ◽  
Healthy Control ◽  
Ifn Γ ◽  
And Control ◽  
Culture Supernatants

Variable intralesional immunotherapies have recently been proposed as a means of achieving a successful eradication of recurrent and recalcitrant human papillomavirus (HPV)-induced cutaneous and anogenital warts. The bivalent HPV vaccine is one of the newly proposed immunotherapeutic agents. We investigated the role of interleukin-4 (IL-4) and interferon-gamma (IFN-γ) as ex vivo immunologic predictors to estimate the response to the bivalent HPV vaccine as a potential immunotherapy for cutaneous and anogenital warts. Heparinized blood samples were withdrawn from forty patients with multiple recurrent recalcitrant cutaneous and anogenital warts and forty matched healthy control subjects. Whole blood cultures were prepared with and without bivalent HPV vaccine stimulation. Culture supernatants were harvested and stored for IL-4 and IFN-γ measurements using an enzyme-linked immunosorbent assay. A comparative analysis of IL-4 and IFN-γ levels in culture supernatants revealed a non-significant change between the patient and control groups. The bivalent HPV vaccine stimulated cultures exhibited a non-significant reduction in IL-4 levels within both groups. IFN-γ was markedly induced in both groups in response to bivalent HPV vaccine stimulation. The bivalent HPV vaccine can give a sensitive IFN-γ immune response ex vivo, superior to IL-4 and sufficient to predict both the successful eradication of HPV infection and the ultimate clearance of cutaneous and anogenital warts when the bivalent HPV vaccine immunotherapy is applied.

scholarly journals Development and applications of a monoclonal antibody against caprine interferon-gamma

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Wen-Tao Ma ◽  
Qi Liu ◽  
Meng-Xia Ning ◽  
Yu-Xu Qi ◽  
Saad Rehman ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Western Blot ◽  
Interferon Gamma ◽  
Mononuclear Cells ◽  
Orf Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Type I ◽  
Fusion Tag ◽  
Wide Range ◽  
Ifn Γ

Abstract Background Interferon-gamma (IFN-γ) is an important mediator of type I immune response and has antiviral, immunoregulatory and anti-tumor properties, plays a wide range of roles in inflammation and autoimmune diseases. The aim of this study was to obtain monoclonal antibody (mAb) against caprine IFN-γ by immunizing of BALB/c mice with the purified rIFN-γ. Results Recombinant caprine IFN-γ was expressed in Escherichia coli strain BL21 (DE3) and monoclonal antibodies against caprine IFN-γ were produced by immunizing of BALB/c mice with rIFN-γ. One hybridoma secreting mAb was screened by enzyme-linked immunosorbent assay (ELISA) which was designated as 2C. MAb secreted by this cell line were analyzed through ELISA, western blot and application of the mAb was evaluated by immunofluorescence analysis using goat lip tissues infected with Orf virus. ELISA analysis revealed that mAb 2C can specifically recognize rIFN-γ protein and culture supernatant of goat peripheral blood mononuclear cells (PBMCs) stimulated by concanavalin A (Con A) but cannot recognize the fusion tag protein of pET-32a. Western blot analysis showed that mAb 2C can specifically react with the purified 34.9 kDa rIFN-γ protein but does not react with the fusion tag protein of pET-32a. Immunofluorescence results demonstrated that mAb 2C can detect IFN-γ secreted in histopathological sites of goats infected with Orf virus. Conclusions A caprine IFN-γ-specific mAb was successfully developed in this study. Further analyses showed that the mAb can be used to detect IFN-γ expression level during contagious ecthyma in goats.

scholarly journals Hypothalamic Glutamine-Glutamate Metabolites and Peripheral Pro-Inflammatory Cytokines in Participants with Depression: A Case-Control Study

2020 ◽  
Author(s):  
Bruno Pedraz Petrozzi ◽  
Carlo Blecker ◽  
Elena Neumann ◽  
Gebhard Sammer
Keyword(s):  
Enzyme Linked Immunosorbent Assay ◽  
Resonance Spectroscopy ◽  
Peripheral Inflammation ◽  
Tnf Α ◽  
Ifn Γ ◽  
And Control ◽  
Pituitary Adrenal Axis ◽  
Inflammatory Component ◽  
Control Study

Abstract Background: In recent years, there has been increasing evidence of an inflammatory component due to overstimulation of the hypothalamic-pituitary-adrenal axis (HPAA) in depression. The glutamate metabolites (glutamate and glutamine) are important in this stimulation. The aim of this study was to determine the concentrations of hypothalamic glutamate metabolites in depression and to investigate their relationship to peripheral inflammation. Methods: Participants with diagnosed depression (DE; n = 24) and control subjects without depression (HC; n = 25) were investigated. Hypothalamic glutamate metabolites were recorded using in vivo magnetic resonance spectroscopy. Peripheral cytokines (IL-6, TNF-α, IFN-γ and IL-1β) were assessed using Enzyme-Linked Immunosorbent Assay. For statistical analysis, generalized mixed models were computed using Poisson distributions and a log link function. Results: The results show overall higher hypothalamic glutamate metabolites in DE compared to HC. High TNF-α and IL-1β concentrations are associated with high hypothalamic glutamate metabolites in DE. Conclusions: These results provide initial evidence that, in depression, increased HPAA activity is associated with peripheral inflammation favored by hypothalamic glutamate metabolites.

Expression profile of interferon-gamma (IFN- γ) mRNA as diagnostic molecular signatures of Hashimoto's Thyroiditis

2021 ◽  
Vol 30 (2) ◽  
pp. 117-123
Author(s):  
Nearmeen M. Rashad ◽  
Reham M. El Shabrawy ◽  
Shereen M. El Shabrawy ◽  
Hassan M. Hassanin
Keyword(s):  
Interferon Gamma ◽  
Hashimoto’S Thyroiditis ◽  
Diagnostic Markers ◽  
Hashimoto's Thyroiditis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Thyroid Function Tests ◽  
Expression Levels ◽  
Egyptian Women ◽  
Ifn Γ ◽  
Obesity Indices

Background: Hashimoto's thyroiditis (HT) is a T cell-mediated autoimmune disease that primarily affects females. IFN- γ is a critical cytokine that has been related to the pathogenesis of HT. Objectives: We aimed to evaluate serum and expression levels of interferon-gamma (IFN- γ) in Egyptian women with HT and to assess the association between serum and expression levels of IFN- γ with clinical and laboratory characteristics of HT. Methodology: This case-control study included 120 women with HT and 70 controls. IFN- γ mRNA expression was analyzed using real-time polymerase chain reaction. Serum IFN- γ was measured using enzyme-linked immunosorbent assay. Results: Serum IFN- γ level and the level of IFN- γ mRNA are both sensitive and specific to be used as diagnostic markers for HT with cut off values of 28.57 pg/ml and 3.55 respectively. Both showed a significant positive correlation with TPO-Ab and Tg-Ab, obesity indices, dyslipidemia, and TSH, while they have a negative correlation with FT3, FT4. Conclusions: Serum IFN- γ level and the level of IFN- γ mRNA are both sensitive and specific to be used as diagnostic markers for HT, significantly correlated with thyroid autoantibodies and thyroid function tests.

Sign in / Sign up

Export Citation Format

Share Document