scholarly journals Toll-Like Receptors 2, 4, and 7, Interferon-Gamma, Interleukin 10, and Programmed Death Ligand 1 Transcripts in Leishmanin Skin Test-Positive Reactions of Ibizan Hound Dogs

2020 ◽  
Vol 2020 ◽  
pp. 1-8
Author(s):  
Laura Ordeix ◽  
Sara Montserrat-Sangrà ◽  
Pamela Martínez-Orellana ◽  
Laia Solano-Gallego
Keyword(s):  
Immune Response ◽  
Skin Test ◽  
Interferon Gamma ◽  
Negative Control ◽  
Immune Gene ◽  
Canine Leishmaniosis ◽  
Healthy Skin ◽  
Gene Expressions ◽  
Leishmanin Skin Test

The leishmanin skin test (LST) is an in vivo technique commonly used to evaluate the Leishmania-specific cellular immune response in dogs. However, information regarding the local immune response in LST-positive reactions is scarce. We examined the pattern of toll-like receptor 2 (TLR2), TLR4, TLR7, interleukin- (IL-) 10, interferon gamma (IFN-γ), and (program death ligand) PD-L1 gene expression in LST-positive reactions and paired normal-looking skin of nine infected Ibizan hound dogs. Healthy skin from ten seronegative dogs from a nonendemic area was analysed as a negative control. Immune gene expressions were examined by quantitative PCR (qPCR) analysis. LST-positive reactions presented significant upregulation of TLR2, TLR4, IL-10, IFN-γ, and PD-L1 and downregulation of TLR7 when compared with healthy skin of seronegative control dogs from a nonendemic area. All transcripts but TLR7 were significantly higher in LST-positive reaction than in paired normal-looking skin of Ibizan hound. The expression profile of immune genes in LST-positive reactions was similar to that previously observed in clinically lesioned skin of mildly diseased dogs with papular dermatitis due to Leishmania infantum infection. This data provide additional support for the important role of TLRs in canine leishmaniosis.

scholarly journals Modulation of Placental Breast Cancer Resistance Protein by HDAC1 in Mice: Implications for Optimization of Pharmacotherapy During Pregnancy

2021 ◽  
Author(s):  
Chuan Wang ◽  
Dan Ma ◽  
Yimin Hua ◽  
Hongyu Duan
Keyword(s):  
Breast Cancer ◽  
Breast Cancer Resistance Protein ◽  
Negative Control ◽  
Great Promise ◽  
Cancer Resistance ◽  
Gene Expressions ◽  
Time Curve ◽  
Resistance Protein

AbstractBreast cancer resistance protein (BCRP/ABCG2) is a critical drug efflux transporters by limiting drugs’ transplacental transfer rates. More investigations on the regulation of placental BCRP offer great promise for enabling pronounced progress in individualized and safe pharmacotherapy during pregnancy. Histone deacetylases (HDACs) play an important role in epigenetic regulation of placental genes. It was reported recently by us that HDAC1 was involved in placental BCRP regulation in vitro. The aim of this study was to further explore the effect of HDAC1 on placental BCRP expression and functionality in animals. Randomly assigned C57BL pregnant dams received intraperitoneal injections of a negative control siRNA or Hdac1 siRNA from embryonic day 7.5 (E7.5) to E15.5, respectively. At E16.5, glyburide (GLB), a probe for evaluating placental BCRP efflux functionality, was injected via the tail vein. Animals were sacrificed through cervical dislocation at various times (5–180 min) after drug administration. The maternal blood, placentas, and fetal-units were collected. GLB concentrations were determined by a validated high-performance liquid chromatography/mass spectrometry (HPLC-MS) assay. Real-time quantitative PCR (qRT-PCR), Western blot, and immunohistochemical (IHC) analysis were employed to identify mRNA/protein levels and localization of gene expressions, respectively. It was noted that Hdac1 inhibition significantly decreased placental Bcrp expression, with markedly increases of GLB concentrations and area under the concentration-time curve (AUC) in fetal-units. Particularly, the ratios of fetal-unit/maternal plasma GLB concentrations were also significantly elevated following Hdac1 repression. Taken together, these findings suggested that HDAC1 was involved in positive regulation of placental BCRP expression and functionality in vivo.

scholarly journals Anti-inflammatory activity of released-active antibodies to interferon-gamma, CD4-receptor, and histamine against respiratory-syncytial viral infection

2019 ◽  
pp. 85-89
Author(s):  
А.Г. Емельянова ◽  
С.А. Тарасов ◽  
С.Г. Морозов
Keyword(s):  
Interferon Gamma ◽  
Inflammatory Cells ◽  
Negative Control ◽  
Ifn Gamma ◽  
Cd4 Receptor ◽  
Cause Of Deaths ◽  
Prophylactic Use ◽  
Syncytial Virus ◽  
One Year

Актуальность. Респираторно-синцитиальный вирус (РСВ) вызывает наиболее опасные инфекции у детей, особенно до 1 года, являясь основной причиной смертельных исходов, и способствует развитию бронхиальной астмы. Эффективной терапии в отношении вызываемой им инфекции не существует, а меры профилактики ограничены. Перспективным может быть использование препаратов на основе релиз-активных антител (РА АТ), действие которых направлено на иммунные реакции организма. Целью работы являлось изучение эффектов РА АТ к ИФН-гамма, CD4-рецептору и гистамину при РСВ-инфекции in vivo при их профилактическом введении. Методы. Мышей линии Balb/c инфицировали интраназально РСВ в дозе 5 × 106 ТЦД50/мышь, в течение 5 суток до инфицирования животным вводили РА АТ к ИФН-гамма, CD4-рецептору и гистамину, или отрицательный контроль (вода очищенная). Через 6 суток после инфицирования оценивали инфильтрацию клеток воспаления в дыхательные пути. Результаты. РА АТ к ИФН-гамма, CD4-рецептору и гистамину статистически значимо (p < 0,05) снижают общую инфильтрацию клеток воспаления в легкие, а также уровень лимфоцитов и макрофагов по сравнению с контрольными группами. Заключение. Профилактическое применение РА АТ к ИФН-гамма, CD4-рецептору и гистамину способствует снижению выраженности воспаления дыхательных путей экспериментальных животных, зараженных РСВ. Background. Respiratory syncytial virus (RSV) causes the most dangerous infections in children, especially those under one year, being the main cause of deaths and contributing to the development of bronchial asthma. There is no effective treatment for the causative infection, and preventive measures are limited. The use of drugs based on released-active antibodies (RA Abs) that target the immune response may be promising. Aim. The aim of the work was to study preventive effects of RA Abs to interferon-gamma (IFN-gamma), CD4 receptor, and histamine on RSV infection in vivo. Methods Balb /c mice were infected with RSV intranasally at a dose of 5 × 106 TCID50 per mouse. For 5 days prior to infection, RA Abs to IFN-gamma, CD4 receptor, and histamine or the negative control (purified water) were intragastrically administered to the animals. Infiltration of inflammatory cells into the respiratory tract was evaluated 6 days after infection. Results. RA Abs to IFN-gamma, CD4 receptor, and histamine significantly (p < 0.05) reduced the total infiltration of inflammatory cells into the lungs, as well as levels of lymphocytes and macrophages compared with the control groups. Conclusion. The prophylactic use of RA Abs to IFN-gamma, CD4 receptor, and histamine helps to alleviate severity of airway inflammation in experimental animals infected with RSV.

scholarly journals Optimizing a Protocol to Assess Immune Responses after SARS-CoV-2 Vaccination in Kidney-Transplanted Patients: In Vivo DTH Cutaneous Test as the Initial Screening Method

Vaccines ◽  
2021 ◽  
Vol 9 (11) ◽  
pp. 1315
Author(s):  
Yvelise Barrios ◽  
Aurelio Rodriguez ◽  
Andrés Franco ◽  
Cristina Alava-Cruz ◽  
Domingo Marrero-Miranda ◽  
...  
Keyword(s):  
Immune Response ◽  
T Cell ◽  
Skin Test ◽  
Natural Infection ◽  
Screening Method ◽  
Delayed Type Hypersensitivity ◽  
Interferon Gamma Release Assay ◽  
Cutaneous Test

Previously, the delayed-type hypersensitivity (DTH) cutaneous test with the spike protein of SARS-CoV-2 has been shown to be a simple in vivo method to measure T-cell functionality after natural infection and in vaccinated individuals. Methods: Twenty-five kidney-transplanted recipients were immunized with two doses of the mRNA-based Pfizer–BioNTech COVID19 vaccine three weeks apart. Cell-immune response (CIR) was evaluated ten weeks later using an in vivo DTH skin test and in vitro with an interferon gamma release assay (IGRA). Humoral Immune Response (HIR) was determined by the measurement of specific IgG anti-S1 SARS-CoV-2. Results: Ten weeks after the second dose of the vaccine, 23 out of 25 transplanted patients had a positive DTH skin test, while in vitro CIR was considered positive in 20 patients. Unspecific stimulation was positive in all 25 patients, showing no T-cell defect. Seven out of twenty-five patients had a negative specific anti-spike IgG. CIR was positive in all immune-competent control patients. Conclusions: DTH is a useful, simple, and cheaper tool that can be used to assess cellular immune response, with an excellent correlation with the in vitro CIR. CIR assessment after vaccination in these immunocompromised patients is an excellent complement to HIR-based methods. This skin test could be used if classical in vitro methods cannot be applied.

scholarly journals Interdigital skin test for evaluation of delayed hypersensitivity and monitoring cell-mediated immune responses in chickens

2007 ◽  
Vol 23 (5-6-2) ◽  
pp. 223-228 ◽  
Author(s):  
B. Miljkovic ◽  
L. Peric ◽  
M. Velhner
Keyword(s):  
Immune Response ◽  
T Cell ◽  
Skin Test ◽  
Skin Testing ◽  
Intradermal Injection ◽  
Delayed Hypersensitivity ◽  
Skin Thickness ◽  
Strong Immune Response ◽  
Cell Mediated Immune Response

A skin test to assess cell mediated delayed hypersensitivity (DH) used to evaluated immune response of chickens. Results of many study indicated, that skin testing is especially useful as a simple in vivo screening to evaluate normal and suppressed T-cell mediated DH. Chickens were sensitized with using mitogens, B and T-cell dependent antigen by intradermal injection. The most feathered skin of chickens is too thin for adequate intradermal injections, so the wattle is the standard site for skin testing, however, in younger than 2 or 3 weeks old chickens, the wattle is undeveloped and intradermal injection and measurement of response are difficult. A simple interdigital skin used by many of the authors. Skin swelling response and DH reaction were measured in mm before injections and after. The skin test and DH in vivo results edemas-initiating characteristics of sensitizing agents, which increase in skin thickness detectable after 4- 6 hours of application. Many of investigation results suggests that healthy chickens are able to have strong immune response and support the concept that some changes in the cell-mediated immune response and other pathogens may potentially affect immune response.

scholarly journals Evaluation of the Immune Response to Interferon Gamma Release Assay and Tuberculin Skin Test Among BCG Vaccinated Children in East of Egypt

Medicine ◽  
2016 ◽  
Vol 95 (17) ◽  
pp. e3470 ◽  
Author(s):  
Mohamed Refaat Beshir ◽  
Alaa Ebrahim Zidan ◽  
Hosam Fathi El-Saadny ◽  
Raghdaa Abdelaziz Ramadan ◽  
Nehad Ahmed Karam ◽  
...  
Keyword(s):  
Immune Response ◽  
Tuberculin Skin Test ◽  
Skin Test ◽  
Interferon Gamma ◽  
Interferon Gamma Release Assay ◽  
Release Assay

scholarly journals A review of the leishmanin skin test: A neglected test for a neglected disease

2021 ◽  
Vol 15 (7) ◽  
pp. e0009531
Author(s):  
Jessica Carstens-Kass ◽  
Kayla Paulini ◽  
Patrick Lypaczewski ◽  
Greg Matlashewski
Keyword(s):  
Skin Test ◽  
Surrogate Marker ◽  
Epidemiological Studies ◽  
Interferon Γ ◽  
Delayed Type Hypersensitivity ◽  
Cell Mediated Immunity ◽  
Leishmanin Skin Test ◽  
Mesh Terms

The leishmanin skin test (LST) has been used for decades to detect exposure and immunity to the parasite Leishmania, the causative agent of the neglected tropical disease leishmaniasis. In the LST, Leishmania antigen (leishmanin) is intradermally injected into the forearm. In an individual who has been previously infected, a delayed-type hypersensitivity (DTH) reaction results in a measurable induration at the site of the injection, indicating that previous exposure to Leishmania has resulted in the development of cell-mediated immunity. LST positivity is associated with long-lasting protective immunity against reinfection, most notably as reported for visceral leishmaniasis (VL). Despite efforts over the past few decades, leishmanin antigen is no longer produced under good manufacturing practice (GMP) conditions anywhere in the world. Consequently, the use of the LST in epidemiological studies has declined in favor of serological and molecular tests. In this review, we provide a historical overview of the LST and justification for the reintroduction of leishmanin. A GMP-grade leishmanin can be used to detect immunity in vivo by the LST and can be investigated for use in an interferon-γ release assay (IGRA), which may serve as an in vitro version of the LST. The LST will be a valuable tool for surveillance and epidemiological studies in support of the VL elimination programs and as a surrogate marker of immunity in vaccine clinical trials. Methods A review of the literature was conducted using PubMed as the primary database, with MeSH terms “leishmanin skin test” OR “Montenegro test” OR “Montenegro skin test.” Articles written in English that describe the history or standardization of leishmanin, the use of leishmanin in an IGRA, or the use of the LST in epidemiological studies or vaccine trials were prioritized in our appraisal of the literature.

scholarly journals FLARIM v2.0, an improved method to quantify transcript-ribosome interactions in vivo in the adult Drosophila brain

2021 ◽  
Author(s):  
Petra Richer ◽  
Sean D Speese ◽  
Mary A Logan
Keyword(s):  
Immune Response ◽  
Morphological Changes ◽  
Immune Gene ◽  
Promising Tool ◽  
Matrix Metalloproteinase 1 ◽  
New Variant ◽  
Wide Range ◽  
Drosophila Brain

Neural injury triggers striking immune reactions from glial cells, including significant transcriptional and morphological changes, but it is unclear how these events are coordinated to mount an effective immune response. Here, we present a new variant of the Fluorescence assay to detect ribosome interactions with mRNA (FLARIM), which we term FLARIM v2.0, to visualize single immune gene transcripts and association with ribosomes in glia responding to neurodegeneration. Specifically, using an in vivo axotomy assay in Drosophila, we show that matrix metalloproteinase-1 (Mmp-1) mRNAs and associated ribosomes are detected in distal processes of reactive glia where they are actively engulfing degenerating axonal material, suggesting that local translation is an important component of the glial immune response to axotomy. This work also validates our enhanced FLARIM assay as a promising tool to investigate mechanisms of mRNA transport and translation in a wide range of in vitro and in vivo paradigms.

scholarly journals Regulatory Network of MiRNAs, IncRNAs, Transcription Factors and Target Genes Associated With Immune Response in Bovine Mastitis

2021 ◽  
Author(s):  
Ashley R. Tucker ◽  
Nicole A. Salazar ◽  
Adeola O. Ayoola ◽  
Erdoğan Memili ◽  
Bolaji N. Thomas ◽  
...  
Keyword(s):  
Immune Response ◽  
Transcription Factors ◽  
Drug Targets ◽  
Target Genes ◽  
Bovine Mastitis ◽  
Regulatory Elements ◽  
Immune Gene ◽  
Pathway Analyses

Abstract Background: Bovine mastitis is a mammary gland infectious disease caused by a variety of pathogens with a devasting economic impact worldwide. Pre- and post-transcriptional modifications, including transcription factors (TFs), altering gene expression are emerging foci of disease studies, with minimal studies revealing the importance of non-coding transcripts, like long non-coding RNAs (lncRNAs) and microRNAs (miRNAs). We hypothesize TFs, lncRNAs and miRNAs can modulate the immune response in bovine mastitis and can potentially serve as disease biomarkers and/or drug targets.Methods: With computational analyses, we aimed to identify candidate bovine mastitis genes and construct the networks of miRNA, lncRNA and TFs regulating the gene’s mRNA, affecting disease pathogenesis. Experimentally validated genes associated with bovine mastitis were obtained through an extensive search for significantly mentioned genes, utilizing several databases. Prediction of miRNA and lncRNA binding bovine mastitis candidate genes were performed through several algorithms and software that relied on base pair complementation, evolutionary conservation, and thermodynamic stability of binding regions. Combined interactome network of lncRNAs, miRNAs, TFs and immune gene targets were constructed.Results: Sixteen of 923 genes were found to be highly significant in bovine mastitis disease pathway including, CD4, IL-10, IFNγ, IL-4, TLR4, TNFα, and CD14. Remarkably, we found six miRNAs, two being bta-miR-223 and bta-miR-24-3p, to bind to several targets. Eight out of 22 lncRNA, such as NONBTAT027932.1 and XR_003029725.1, were identified as regulatory elements that target the genes based on the normalized binding free energies ranging from -0.1774 to -2.875. Similarly identified were 17 TFs, including JUN and CREB. Our functional and pathway analyses revealed several pathways like lipopolysaccharide-mediated signaling pathway, regulation of chemokine (C-X-C motif) ligand 2 production and regulation of IL-23 production among others. Conclusions: The overarching interactome in this study is the first of its kind regarding bovine mastitis, deserving further in vitro/in vivo explication for specific molecular regulatory mechanisms during bovine mastitis immune response, which could lay the foundation for development of disease markers and therapeutic intervention.

scholarly journals Development of Cutaneous Leishmaniasis after Leishmania Skin Test

2011 ◽  
Vol 2011 ◽  
pp. 1-4 ◽  
Author(s):  
Paulo R. Machado ◽  
Augusto M. Carvalho ◽  
Gustavo U. Machado ◽  
Marina L. Dantas ◽  
Sérgio Arruda
Keyword(s):  
Immune Response ◽  
Cutaneous Leishmaniasis ◽  
Skin Test ◽  
Mononuclear Cells ◽  
Previous History ◽  
Self Healing ◽  
Necrosis Factor Alpha ◽  
Cutaneous Ulcer ◽  
The Right

Thirty-year-old female with a previous history of a cutaneous ulcer suspicious of leishmaniasis 20 years ago presented with a new complaint of a depressed papular lesion8×7 mm in the right lower extremity. The lesion was of 10-day duration. Because early cutaneous leishmaniasis (CL) lesions may have a non-ulcerated appearance, a Leishmania skin test (LST) was performed on the forearm with a strong positive result (38×32 mm). After 8 days, the lesion in the leg, which was diagnosed as folliculitis, completely healed. However, a typical CL ulcer (26×24 mm) developed at the LST site. Histopathology of the new lesion did not identifiy parasites, but the findings were consistent with a diagnosis of CL. Further analysis identified amastigotes by immunohistochemical stain. Mononuclear cells harvested from the patient were stimulated with Leishmania antigen and showed high levels of production of both tumor necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ): 2,943 pg/mL and 2,313 pg/mL, respectively. After 40 days of treatment with antimony and pentoxifylline, the ulcer resolved. The development of CL at the LST site suggests a strong Th1 immune response, and it is anin vivodocumentation of the role of the host immune response in the pathology of CL. It teaches us that LST should be cautiously, if at all, used in patients with self-healing CL ulcers.

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