Local Cellular Immune Response Research Articles

The local skin cellular immune response determines the clinical outcome of sarcoptic mange in Iberian ibex (Capra pyrenaica).

Sarcoptic mange, caused by Sarcoptes scabiei, is a disease with implications for wildlife conservation and management. Its severity depends on the host's local skin immune response, which is largely unknown in Iberian ibex (Capra pyrenaica), a mountain ungulate dramatically affected by mange. In this species, the clinical outcome of sarcoptic mange varies among individuals, and the local immune response could be key to controlling the infestation. This study aims to characterize the local cellular immune response and its relationship with the clinical outcome. Fourteen Iberian ibexes were experimentally infested with S. scabiei and six more served as controls. Clinical signs were monitored, and skin biopsies were collected from the withers at 26, 46, and 103 days post-infection (dpi). The presence and distribution of macrophages (including M1 and M2 phenotypes), T lymphocytes, B lymphocytes, plasma cells, and interleukine 10 were quantitatively evaluated using immunohistochemical techniques. An inflammatory infiltrate that decreased significantly from 26 to 103 dpi was observed in all the infested ibexes. The predominant inflammatory cell population in the skin of the mangy ibexes was formed by macrophages (mainly the M2 phenotype) followed by T lymphocytes, with lower numbers of B lymphocytes and plasma cells. Three clinical courses were identified: total recovery, partial recovery, and terminal stage. The inflammatory infiltrates were less pronounced in the fully recovered ibexes than in those that progressed to the terminal stage throughout the study. The results suggest an exacerbated but effective Th1-type cellular immune response controlling mange in Iberian ibex. Furthermore, the local immune response appears to determine the variability of the clinical responses to S. scabiei infestation in this species. This first report on the progression of local skin immune cells is relevant not only for individuals but also for population management and conservation.

The Role of Dietary Fibre in Enteral Nutrition in Sepsis Prevention and Therapy: A Narrative Review.

This narrative review summarises the current evidence on the role of dietary fibre in enteral nutrition in the prevention and therapy of sepsis, with a focus on critically ill patients. The aim is to discuss the implications for clinical practice and identify future directions for policy and research. We searched MEDLINE and Google Scholar for records on sepsis, critically ill, enteral nutrition, and dietary fibre. We included all types of articles such as meta-analyses, reviews, clinical trials, preclinical studies, and in vitro studies. Data were evaluated for significance and clinical relevance. Synopsis of Review: Despite the ongoing debate, enteral nutrition containing dietary fibres showed great potential in attenuating sepsis-related outcomes and preventing the incidence of sepsis in critically ill patients on enteral nutrition. Dietary fibres target different underlying mechanisms such as microbiota, mucosal barrier integrity, local cellular immune response, and systemic inflammation. We discuss the clinical potential and concerns that currently exist with the standard implementation of dietary fibre in enterally fed intensive care patients. Additionally, we identified research gaps that should be addressed to determine effectiveness and the role of dietary fibres in sepsis itself and its associated outcomes.

Local cellular immune response plays a key role in protecting chickens against hepatitis-hydropericardium syndrome (HHS) by vaccination with a recombinant fowl adenovirus (FAdV) chimeric fiber protein.

Fowl adenovirus (FAdV)-induced diseases hepatitis-hydropericardium syndrome (HHS) and inclusion body hepatitis (IBH) have been affecting the poultry industry with increasing severity in the last two decades. Recently, a subunit vaccine based on a chimeric fiber protein with epitopes from different fowl adenovirus serotypes (named crecFib-4/11) has been shown to confer simultaneous protection against both HHS and IBH. However, the underlying immune mechanisms in chickens are still enigmatic, especially because of frequently absent neutralizing response despite high levels of protection. In this study, we investigated the kinetics of the humoral and cellular immune responses in specific pathogen-free chickens after vaccination with crecFib-4/11 and/or challenge with a HHS-causing strain, on a systemic level, as well as locally in target and lymphoid organs. The humoral response was assessed via enzyme-linked immunosorbent assay (ELISA) and virus neutralization test in serum, while the cellular immune response was determined by phenotyping using flow cytometry. Although vaccination induced serum antibodies, as confirmed by ELISA, such antibodies exhibited no pre-challenge neutralizing activity against FAdV-4. Nevertheless, immunized birds experienced a significant B cell increase in the liver upon challenge, remaining high throughout the experiment. Furthermore, vaccination stimulated the proliferation of cytotoxic T lymphocytes, with earlier circulation in the blood compared to the challenge control and subsequent increase in liver and spleen. Overall, these findings imply that protection of chickens from HHS after crecFib-4/11 vaccination relies on a prominent local immune response in the target organs, instead of circulating neutralizing antibodies.

Polycationic HA/CpG Nanoparticles Induce Cross-Protective Influenza Immunity in Mice.

The intranasal (i.n.) route is an ideal vaccination approach for infectious respiratory diseases like influenza. Polycationic polyethylenimine (PEI) could form nanoscale complexes with negatively charged viral glycoproteins. Here we fabricated PEI-hemagglutinin (HA) and PEI-HA/CpG nanoparticles and investigated their immune responses and protective efficacies with an i.n. vaccination regimen in mice. Our results revealed that the nanoparticles significantly enhanced HA immunogenicity, providing heterologous cross-protection. The conserved HA stalk region induced substantial antibodies in the nanoparticle immunization groups. In contrast to the Th2-biased, IgG1-dominant antibody response generated by PEI-HA nanoparticles, PEI-HA/CpG nanoparticles generated more robust and balanced IgG1/IgG2a antibody responses with augmented neutralization activity and Fc-mediated antibody-dependent cellular cytotoxicity (ADCC). PEI-HA/CpG nanoparticles also induced enhanced local and systemic cellular immune responses. These immune responses did not decay over six months of observation postimmunization. PEI and CpG synergized these comprehensive immune responses. Thus, the PEI-HA/CpG nanoparticle is a potential cross-protective influenza vaccine candidate. Polycationic PEI nanoplatforms merit future development into mucosal vaccine systems.

Immunophenotype of the inflammatory response in the central and enteric nervous systems of cockatiels (Nymphicus hollandicus) experimentally infected with parrot bornavirus 2.

Proventricular dilatation disease is a lethal disease of psittacine birds. In this study, we characterized the local cellular immune response in the brain, proventriculus, and small intestine of 27 cockatiels (Nymphicus hollandicus) experimentally infected with parrot bornavirus 2 (PaBV-2). Perivascular cuffs in the brain were composed of CD3+ T-lymphocytes and Iba1+ macrophages/microglia in most cockatiels (n = 26). In the ganglia of the proventriculus, CD3+ T-lymphocytes (n = 17) and Iba1+ macrophages (n = 13) prevailed. The ganglia of the small intestine had a more homogeneous distribution of these leukocytes, including PAX5+ B-lymphocytes (n = 9), CD3+ T-lymphocytes (n = 8), and Iba1+ macrophages (n = 8). Our results indicate that perivascular cuffs in the brain and the inflammatory infiltrate in the proventriculus of PaBV-2-infected cockatiels is predominately composed of T-lymphocytes, while the inflammatory infiltrates in the ganglia of the small intestine are characterized by a mixed infiltrate composed of T-lymphocytes, B-lymphocytes, and macrophages.

Local cellular immune response in chronic periodontitis

Microflora of the oral cavity forms a biofilm that induces response of immune system at the mucous membranes. Transition to periodontal lesion is provided by certain classes of resident mucosal immune cells and inflammatory/immune cells migrating to the periodont. In periodontal diseases, Th1, Th2, Th17, Treg are detected. T regulatory cells (Tregs) are proven to comprise the main anti-inflammatory cell population. Th17 cells and Treg cells play an important role in osteoclast differentiation. IL-17 secreted by Th17 cells affects osteoclastogenesis and may induce macrophages to enhance the local inflammatory response. In this regard, the aim of our work was to identify the local immune cells in oral cavity which are associated with severity of chronic generalized periodontitis. The oral cavity cells from 58 persons aged 38-65 years of both sexes in their mature age with a diagnosis of «chronic periodontitis» were examined by means of flow cytofluorometry. When determining levels of CD64+CD16+CD14- neutrophils in the patients with periodontitis of different severity, a statistically significant increase of this cell population was revealed upon development of this disease. In mild cases of periodontitis, a significant increase of relative CD64+CD16+CD14- neutrophil contents was revealed (Me = 36.16%, p < 0.05) compared to the control group (Me = 7.7%, Q0.25 = 2.4%, Q0.75 = 12%). When assessing relative numbers of CD14+ monocytes in periodontitis of various severity, we revealed a significant increase in the number of these cells in severe cases. When studying levels of regulatory T lymphocytes (CD4+CD25+CD127low) in periodontitis of different severity, we revealed significantly decreased amounts of this cell population during development of the disease. In mild cases of periodontitis, a decreased level of CD4+CD25+CD127low cells (p < 0.05, Me = 1356 cells/ml) was revealed, as compared with control group (Me = 10666 cells/ml). Although the concentration of CD4+CD25+CD127low (Me = 4709 cells/ml) in the patients with moderate periodontitis was higher than the values in milder cases, the range of the main values was comparable and lower, than in control group. In severe periodontitis, a significantly decreased concentration of regulatory T lymphocytes was revealed (Me = 2637 cells/ml). These results confirm the anti-inflammatory regulatory function of Tregs. Understanding the osteo-immune mechanisms of bone remodeling control will help to understand the pathophysiology of accelerated bone loss observed in severe chronic periodontitis.

The Dietary Intake of Carrot-Derived Rhamnogalacturonan-I Accelerates and Augments the Innate Immune and Anti-Viral Interferon Response to Rhinovirus Infection and Reduces Duration and Severity of Symptoms in Humans in a Randomized Trial.

Acute respiratory infections are an important health concern. Traditionally, polysaccharide-enriched extracts from plants, containing immunomodulatory rhamnogalacturonan-I (RG-1), were used prophylactically. We established the effects of dietary supplementation with carrot-derived RG-I (cRG-I, 0–0.3–1.5 g/day) in 177 healthy individuals (18–65 years) on symptoms following infection with rhinovirus strain 16 (RV16). Primary outcomes were changes in severity and duration of symptoms, and viral load in nasal lavage. Secondary outcomes were changes in innate immune and anti-viral responses, reflected by CXCL10 and CXCL8 levels and cell differentials in nasal lavage. In a nested cohort, exploratory transcriptome analysis was conducted on nasal epithelium. Intake of cRG-I was safe, well-tolerated and accelerated local cellular and humoral innate immune responses induced by RV16 infection, with the strongest effects at 1.5 g/d. At 0.3 g/d, a faster interferon-induced response, induction of the key anti-viral gene EIF2AK2, faster viral clearance, and reduced symptom severity (−20%) and duration (−25%) were observed. Anti-viral responses, viral clearance and symptom scores at 1.5 g/d were in between those of 0 and 0.3 g/d, suggesting a negative feedback loop preventing excessive interferon responses. Dietary intake of cRG-I accelerated innate immune and antiviral responses, and reduced symptoms of an acute respiratory viral infection.

Clinical and immunological characteristics of tegumentary leishmaniasis cases in Bolivia.

BackgroundTegumentary leishmaniasis (TL) is a parasitic disease that can present a cutaneous or mucocutaneous clinical form (CL and MCL, respectively). The disease is caused by different Leishmania species and transmitted by phlebotomine sand flies. Bolivia has one of the highest incidences of the disease in South America and the diagnosis is done by parasitological techniques. Our aim was to describe the clinical and immunological characteristics of CL and MCL patients attending the leishmaniasis reference center in Cochabamba, Bolivia, in order to gain updated clinical and epidemiological information, to evaluate the diagnostic methods used and to identify biomarkers related to clinical disease and its evolution.Methodology/Principal findingsThe study was conducted from September 2014 to November 2015 and 135 patients with lesions compatible with CL or MCL were included. Epidemiological and clinical data were collected using a semi-structured questionnaire. Two parasitological diagnostic methods were used: Giemsa-stained smears and culture of lesion aspirates. Blood samples obtained from participants were used to measure the concentrations of different cytokines. 59.2% (80/135) were leishmaniasis confirmed cases (CL: 71.3%; MCL: 28.7%). Sixty percent of the confirmed cases were positive by smears and 90.6% were positive by culture. 53.8% were primo-infections. Eotaxin and monokine induced by IFN-γ presented higher serum concentrations in the MCL clinical presentation compared to CL cases and no-cases. None of the cytokines presented different concentrations between primo-infections and secondary infections due to treatment failure.Conclusions/SignificanceIn Bolivia, parasitological diagnosis remains the reference standard in diagnosis of leishmaniasis because of its high specificity, whereas the sensitivity varies over a wide range leading to loss of cases. Until more accurate tools are implemented, all patients should be tested by both smears and culture of lesion aspirates to minimize the risk of false negatives. Our results showed higher concentrations of several cytokines in MCL compared to CL, but no differences were observed between CL and no-cases. In addition, none of the cytokines differed between primary and secondary infections. These results highlight the need of further research to identify biomarkers of susceptibility and disease progression, in addition to looking at the local cellular immune responses in the lesions.

Research Progress of HPV L1 Capsid Protein in Prediction of Cervical Lesions

Cervical cancer is one of the most common malignant gynecological tumors and has the second highest incidence of all malignancies in females. Chronic and persistent infection with High Risk Human Papillomavirus (HR-HPV) is the main cause of cervical cancer. There is a distinct lack of methodology by which to determine whether cervical epithelial dysplasia is cancerous following HPV infection. HPV L1 capsid protein is a major structural protein of human papillomavirus (HPV)?and it is the main target of the local cellular immune response aiming to combat human papillomavirus after HPV infection within cervical cells. Greater understanding of HPV L1 capsid protein and its association with cervical cytology, histopathology, patient age and human papillomavirus viral load has the potential to contribute toward improved the improved diagnosis and management of cervical cancer, providing useful information for gynecological clinicians in the hope of improving patient treatment and quality of life. This article reviews the predictive utility of HPV L1 capsid protein for cervical lesions.

Establishing a Multicolor Flow Cytometry to Characterize Cellular Immune Response in Chickens Following H7N9 Avian Influenza Virus Infection.

Avian influenza virus (AIV) emerged and has continued to re-emerge, continuously posing great threats to animal and human health. The detection of hemagglutination inhibition (HI) or virus neutralization antibodies (NA) is essential for assessing immune protection against AIV. However, the HI/NA-independent immune protection is constantly observed in vaccines’ development against H7N9 subtype AIV and other subtypes in chickens and mammals, necessitating the analysis of the cellular immune response. Here, we established a multi-parameter flow cytometry to examine the innate and adaptive cellular immune responses in chickens after intranasal infection with low pathogenicity H7N9 AIV. This assay allowed us to comprehensively define chicken macrophages, dendritic cells, and their MHC-II expression, NK cells, γδ T cells, B cells, and distinct T cell subsets in steady state and during infection. We found that NK cells and KUL01+ cells significantly increased after H7N9 infection, especially in the lung, and the KUL01+ cells upregulated MHC-II and CD11c expression. Additionally, the percentages and numbers of γδ T cells and CD8 T cells significantly increased and exhibited an activated phenotype with significant upregulation of CD25 expression in the lung but not in the spleen and blood. Furthermore, B cells showed increased in the lung but decreased in the blood and spleen in terms of the percentages or/and numbers, suggesting these cells may be recruited from the periphery after H7N9 infection. Our study firstly disclosed that H7N9 infection induced local and systemic cellular immune responses in chickens, the natural host of AIV, and that the flow cytometric assay developed in this study is useful for analyzing the cellular immune responses to AIVs and other avian infectious diseases and defining the correlates of immune protection.

Molecular characterization of a novel aspartyl protease-1 from Trichinella spiralis

The aim of this study was to characterize the biological properties of a novel aspartic protease-1 from Trichinella spiralis (TsASP1) and evaluate its potential in inducing immune response. TsASP1 gene was cloned and expressed in Escherichia coli BL21 (DE3). On Western blotting analysis with anti-rTsASP1 serum, native TsASP1 was detected in various T. spiralis phases other than newborn larvae (NBL). qPCR results showed that TsASP1 transcription was the highest in intestinal infective larvae (IIL) and the lowest in the NBL stage. Immunofluorescence test result shows that native TsASP1 was principally localized in stichosome, muscle cells of muscle larvae (ML) and IIL, and surrounded intrauterine embryos in female adult worms (AW). After silencing TsASP1 gene of the ML by siRNA, the worm development was significantly inhibited, showed by shorter AW and more wrinkles and longitudinal crack on epicuticle of AW on scanning electron microscopy; the AW and ML burdens were reduced by 41.82 and 56.36% respectively, compared with the control siRNA or PBS group (P < 0.001). Immunization of mice with rTsASP1 elicited an evident antibody response (serum IgG, IgG1/IgG2a and enteral sIgA), and systemic (spleen) and intestinal local mucosal (mesenteric lymph node) cellular immune response, demonstrated by a prominent elevation of IFN-γ and IL-4. The results suggested TsASP1 participated in T. spiralis development and survival in host, and immunization of mice with rTsASP1 induced systemic/intestinal local mucosal humoral and cellular immune response against Trichinella.

Vaccination of Mice with a Novel Trypsin from Trichinella spiralis Elicits the Immune Protection against Larval Challenge.

Trichinella spiralis is a major foodborne parasite and has a serious threat to meat safety. Development of anti-Trichinella vaccines is prospective to eliminate Trichinella infection in food animal. The aim of this study was to assess the biological properties of a novel T. spiralis trypsin (TsT) and its elicited immune protection against larval challenge. The cDNA sequence of TsT gene was cloned and expressed. Western blotting showed rTsT was identified by infection serum and anti-TsT serum. RT-PCR results revealed that TsT gene was transcribed at diverse T. spiralis lifecycle stages. The IIFT results showed that natural TsT was principally expressed at epicuticle of 5-6 day adult worms, indicating that TsT is a worm somatic antigen and adult-stage specific surface antigen. Vaccination of mice with rTsT triggered an evident humoral immune response (high levels of serum IgG, IgG1/IgG2a, and enteral sIgA), and it also induced the systemic and enteral local cellular immune response, demonstrated by an significantly elevation of cytokines IFN-γ and IL-4. The mice vaccinated with rTsT exhibited a 33.17% reduction of enteral adult worms and a 37.80% reduction of muscle larvae after larval challenge. The results showed that TsT might be considered as a candidate target antigen for anti-T. spiralis vaccines.

Cells and Cytokines in Milk of Subclinically Infected Bovine Mammary Glands after the Use of Immunomodulatory Composition GLP 810.

The aim of this study was to investigate the effect of intramammary infusions of natural composition GLP 810 with immunomodulating properties on the local nonspecific cellular and humoral immune response in cows with subclinical mastitis. The composition GLP 810 consists of lactic acid, lysozyme, glycopeptides, and 0.9% solution of NaCl. The following parameters were studied: (1) leukocyte differential distribution in milk, (2) expression of cytokines in milk leukocytes, (3) antibacterial activity, and (4) milk quality. Nineteen mammary glands in five lactating cows were infused with 10 mL of GLP 810, and nineteen other glands from five control cows were treated with 10 mL 0.9% NaCl. The results showed that after intramammary administration of the composition GLP 810 three times with 48 h intervals, the following effects on leukocyte populations in milk were observed: (1) an increase in the number of polymorphonuclear leukocytes and lymphocytes and (2) a decrease in the number of macrophages. A reduction in the number of pathogenic bacteria was also detected. The analyses of tumour necrosis factor-alpha, interleukin-10, and beta-defensin-2 revealed that the production of the aforementioned cytokines significantly increased, whereas no significant effects on interleukin-1 and caspase-6 expression in milk leukocytes were recorded. However, there were significant differences between mammary glands with high and low milk somatic cell count. The results suggest that the composition GLP 810 has an immunomodulatory effect on mammary glands and it could be used for improving the immune response in cows with subclinical mastitis during lactation.

A CYTOLOGICAL STUDY OF IMPRINT SMEARS (TOUCH PREPARATION CYTOLOGY) TO EVALUATE THE TOXICITY OF METAL-CONTAINING NANOPARTICLES IN EXPERIMENTAL ANIMALS

Introduction. Touch preparation cytology is a well-known technique widely used in clinical practice. It can be also applied for an express assessment of cyto-morphological effects of metal-containing nanoparticles in experimental animals. Material and methods. We’ve studied 144 imprint smears (of the liver, kidneys, lungs, tracheobronchial and mesenteric lymph nodes) taken from 52 rats, weighed 280-300 g, aged 3.5 months. This was done following a subchronic intraperitoneal administration of TiO2, Al2O3, and SiO2 nanoparticles, in a range of doses, and a subacute (5-times) inhalational exposure to 1 mg/m3 NiO NPs. Leishman stain was used. Cytological markers were studied at х100/х1000 magnification, using a Carl Zeiss Primo Star microscope, with a USCMOS camera for visualization. The count was done for every 200 cells of lung, liver and kidney specimens and for every 100 cells of lymph node tissue. The differences between the group means (in case of quantitative results) were analyzed using a Student’s t-test. The differences between the group means were considered statistically significant if the probability of those differences being accidental didn’t exceed 5% (р &lt; 0,05). Results. We report the principal results obtained in a cytological study of imprint smears (touch preparations) of rat organs following the exposure to metal-containing nanoparticles. The main types of cellular responses were determined. Degenerative and dystrophic changes were observed in touch preparations of the liver, kidneys, and lungs following administration of nanoparticles in a range of doses and by various routes of exposure. Hyperergic inflammatory responses were seen upon the exposure to nickel oxide nanoparticles. A local cellular immune response was observed as an increase of macrophage percentage in imprint smears of some organs. Conclusion. The use of touch preparation cytology together with histological examination of tissue specimens expand opportunities for a more relevant morphological and quantitative analysis when studying the toxic effects of metal-containing nanoparticles.

ПРИМЕНЕНИЕ ЦИТОЛОГИЧЕСКОГО МЕТОДА ИССЛЕДОВАНИЯ МАЗКОВ У ЭКСПЕРИМЕНТАЛЬНЫХ ЖИВОТНЫХ ДЛЯ ОЦЕНКИ ТОКСИЧЕСКОГО ДЕЙСТВИЯ МЕТАЛЛОСОДЕРЖАЩИХ НАНОЧАСТИЦ

Introduction. Touch preparation cytology is a well-known technique widely used in clinical practice. It can be also applied for an express assessment of cyto-morphological effects of metal-containing nanoparticles in experimental animals. Material and methods. We’ve studied 144 imprint smears (of the liver, kidneys, lungs, tracheobronchial and mesenteric lymph nodes) taken from 52 rats, weighed 280-300 g, aged 3.5 months. This was done following a subchronic intraperitoneal administration of TiO2, Al2O3, and SiO2 nanoparticles, in a range of doses, and a subacute (5-times) inhalational exposure to 1 mg/m 3 NiO NPs. Leishman stain was used. Cytological markers were studied at х100/х1000 magnification, using a Carl Zeiss Primo Star microscope, with a USCMOS camera for visualization. The count was done for every 200 cells of lung, liver and kidney specimens and for every 100 cells of lymph node tissue. The differences between the group means (in case of quantitative results) were analyzed using a Student’s t-test. The differences between the group means were considered statistically significant if the probability of those differences being accidental didn’t exceed 5% (р < 0,05). Results. We report the principal results obtained in a cytological study of imprint smears (touch preparations) of rat organs following the exposure to metal-containing nanoparticles. The main types of cellular responses were determined. Degenerative and dystrophic changes were observed in touch preparations of the liver, kidneys, and lungs following administration of nanoparticles in a range of doses and by various routes of exposure. Hyperergic inflammatory responses were seen upon the exposure to nickel oxide nanoparticles. A local cellular immune response was observed as an increase of macrophage percentage in imprint smears of some organs. Conclusion. The use of touch preparation cytology together with histological examination of tissue specimens expand opportunities for a more relevant morphological and quantitative analysis when studying the toxic effects of metal-containing nanoparticles.

Detection of Antigen-Specific T Cells Using In Situ MHC Tetramer Staining.

The development of in situ major histocompatibility complex (MHC) tetramer (IST) staining to detect antigen (Ag)-specific T cells in tissues has radically revolutionized our knowledge of the local cellular immune response to viral and bacterial infections, cancers, and autoimmunity. IST combined with immunohistochemistry (IHC) enables determination of the location, abundance, and phenotype of T cells, as well as the characterization of Ag-specific T cells in a 3-dimensional space with respect to neighboring cells and specific tissue locations. In this review, we discuss the history of the development of IST combined with IHC. We describe various methods used for IST staining, including direct and indirect IST and IST performed on fresh, lightly fixed, frozen, and fresh then frozen tissue. We also describe current applications for IST in viral and bacterial infections, cancer, and autoimmunity. IST combined with IHC provides a valuable tool for studying and tracking the Ag-specific T cell immune response in tissues.

The Proteomic Effects of Pulsed Focused Ultrasound on Tumor Microenvironments of Murine Melanoma and Breast Cancer Models

Non-ablative pulsed focused ultrasound (pFUS) targets non-thermal forces that activate local molecular and cellular immune responses. Optimal parameters to stimulate immunotherapeutic tumor microenvironments (TME) and responses in different tumor types remain uninvestigated. Flank B16 murine melanoma and 4T1 breast tumors received 1 MHz pFUS at 1–8 MPa peak negative pressures (PNP) and were analyzed 24 hr post-sonication. Necrosis or hemorrhage were unaltered in both tumors, but pFUS induced DNA strand breaks in tumor cells at PNP ≥6 MPa. pFUS at >4 MPa suppressed anti-inflammatory cytokines in B16 tumors. pFUS to 4T1 tumors decreased anti-inflammatory cytokines and increased pro-inflammatory cytokines and cell adhesion molecules. pFUS at 6 MPa increased calreticulin and alterations in check-point proteins along with tumoral and splenic immune cell changes that could be consistent with a shift towards an anti-TME. pFUS-induced TME alterations shows promise in generating anti-tumor immune responses, but non-uniform responses between tumor types require additional investigation to assess pFUS as a suitable anti-tumor therapy.

Detection of Humoral and Cellular Immune Responses in Buffaloes Naturally Infected with Sarcocystosis with Risk Factor Assessment

AbstractSarcocystosis is a silent, parasitic disease which affects various animal species and causes significant economic losses. It is caused by a number of different intracellularSarcocystisspp. This study was aimed to detect the host humoral and cellular immune response due to natural infection. Adding to the determination of the infection rate in Monufia Governorate, Egypt. A total number of 127 Egyptian buffaloes (Bubalus bubalis); 30 males and 97 females between 2-11 years of age were examined during 2018. An infection rate of 74% (94/127) was detected by macroscopic examination. The old age females were found to be at a high risk of 90.7% (88/97) in comparison with the young males 20% (6/30). Immunologically, the cellular and humoral immune response was determined using ELISA. A marked down-regulation of the proinflammatory Th-1 cytokine (IFN-γ) and up-regulation of the anti-inflammatory Th-2 cytokine (IL-5) adding to a high level of IgG and IgE were detected in the infected animals compared to the non infected ones. The local cellular immune response in the infected tissues was characterized by an accumulation of mixed inflammatory cells, granuloma formation, eosinophilic infiltration, muscular edema, and necrotic degeneration. In conclusion, theSarcocystisinfection rate in the naturally infected buffaloes in Monufia Governorate was high. This is the first study to provide a fundamental insight into the immune profile in buffaloes infected withSarcocystisspp. So, it will provide valuable insights to develop novel effective vaccines in future studies. Moreover, sensitive and specific tools should be established for the accurate diagnosis of this disease in the different Egyptian governorates through well-structured serological surveys.

Innovative Mucosal Vaccine Formulations Against Influenza A Virus Infections.

Despite efforts made to develop efficient preventive strategies, infections with influenza A viruses (IAV) continue to cause serious clinical and economic problems. Current licensed human vaccines are mainly inactivated whole virus particles or split-virion administered via the parenteral route. These vaccines provide incomplete protection against IAV in high-risk groups and are poorly/not effective against the constant antigenic drift/shift occurring in circulating strains. Advances in mucosal vaccinology and in the understanding of the protective anti-influenza immune mechanisms suggest that intranasal immunization is a promising strategy to fight against IAV. To date, human mucosal anti-influenza vaccines consist of live attenuated strains administered intranasally, which elicit higher local humoral and cellular immune responses than conventional parenteral vaccines. However, because of inconsistent protective efficacy and safety concerns regarding the use of live viral strains, new vaccine candidates are urgently needed. To prime and induce potent and long-lived protective immune responses, mucosal vaccine formulations need to ensure the immunoavailability and the immunostimulating capacity of the vaccine antigen(s) at the mucosal surfaces, while being minimally reactogenic/toxic. The purpose of this review is to compile innovative delivery/adjuvant systems tested for intranasal administration of inactivated influenza vaccines, including micro/nanosized particulate carriers such as lipid-based particles, virus-like particles and polymers associated or not with immunopotentiatory molecules including microorganism-derived toxins, Toll-like receptor ligands and cytokines. The capacity of these vaccines to trigger specific mucosal and systemic humoral and cellular responses against IAV and their (cross)-protective potential are considered.

Immunological Aspects of Trichomonas vaginalis Infection in Women Attending Maternity Teaching Hospital and Some Public Health Centers in Erbil Governorate, Northern Iraq

Trichomonas vaginalis infection is one of the sexually transmitted diseases. It is a health problem all over the world, including Iraq. It has also been associated with adverse outcomes of pregnancy and increased risk of HIV. Trichomoniasis typically elicits aggressive local cellular immune responses with inflammation of the vaginal epithelium and exocervix in women and urethra in men. The present study was aimed to assess serum level of interferon gamma (IFN-γ), Interleukin 10 (IL-10), C-reactive protein, antiphospholipid, anticardiolipin antibodies with eosinophil count as well. Four hundred and forty women with ages ranging between 16 and 60 years old (average of 34.2 years) who attended Maternity Teaching Hospital and a number of public health centers in Erbil Governorate were screened for trichomoniasis by direct wet mount preparation and culture technique. Serum IFN-γ, IL-10, antiphospholipid, and anticardiolipin were assessed by enzyme-linked immunosorbent assay. C-reactive protein (CRP) and eosinophil count were assessed by I CROMA and Coulter, respectively. The results revealed that IL-10 level (96.46 ± 1.97 pg/ml vs. 91.86 ± 1.48 pg/ml), eosinophil count (0.1 ± 0.07 × 10³/μL vs. 0.1 ± 0.01 × 10³/μL), and CRP concentration (2.55 ± 0.74 mg/l vs. 2.27 ± 0.37 mg/l) were non-significantly (P &gt; 0.05) changed in infected women in comparison with negative control group. However, serum IFN-γ level (484.83 ± 38.35 pg/ml vs. 372.15 ± 9.49 pg/ml) was significantly (P &lt; 0.05) elevated in infected women in comparison to the control group. The results also revealed significant increase (P &lt; 0.05) of antiphospholipid immunoglobulin G (IgG) antibodies (4.75 ± 0.3 U/ml vs. 3.79 ± 0.11 U/ml) in the sera of infected women, but antiphospholipid immunoglobulin M (IgM), anticardiolipin IgG, and anticardiolipin IgM antibodies were non-significantly altered in response to T. vaginalis infection.