Mediators Of Cellular Immunity Research Articles

Pathogenicity and escape to pre-existing immunity of a new genotype of swine influenza H1N2 virus that emerged in France in 2020

In 2020, a new genotype of swine H1N2 influenza virus (H1avN2–HA 1C.2.4) was identified in France. It rapidly spread within the pig population and supplanted the previously predominant H1avN1-HA 1C.2.1 virus. To characterize this new genotype which is genetically and antigenically distant from the other H1avNx viruses detected in France, an experimental study was conducted to compare the outcomes of H1avN2 and H1avN1 infections in pigs and evaluate the protection conferred by the only inactivated vaccine currently licensed in Europe containing an HA 1C (clade 1C.2.2) antigen. Infection with H1avN2 induced stronger clinical signs and earlier shedding than H1avN1. The neutralizing antibodies produced following H1avN2 infection were unable to neutralize H1avN1, and vice versa, whereas the cellular-mediated immunity cross-reacted. Vaccination slightly altered the impact of H1avN2 infection at the clinical level, but did not prevent shedding of infectious virus particles. It induced a cellular-mediated immune response towards H1avN2, but did not produce neutralizing antibodies against this virus. As in vaccinated animals, animals previously infected by H1avN1 developed a cross-reacting cellular immune response but no neutralizing antibodies against H1avN2. However, H1avN1 pre-infection induced a better protection against the H1avN2 infection than vaccination, probably due to higher levels of non-neutralizing antibodies and a mucosal immunity. Altogether, these results showed that the new H1avN2 genotype induced a severe respiratory infection and that the actual vaccine was less effective against this H1avN2-HA 1C.2.4 than against H1avN1-HA 1C.2.1, which may have contributed to the H1avN2 epizootic and dissemination in pig farms in France.

Plasma immune signatures can predict rejection-free survival in the first year after pediatric liver transplantation

After pediatric liver transplantation (pLT), children undergo life-long immunosuppression since reliable biomarkers for the assessment of rejection probability are scarce. In the multicentre (n=7) prospective clinical cohort "ChilSFree" study, we aimed to characterize longitudinal dynamics of soluble and cellular immune mediators during the first year after pLT and identify early biomarkers associated with outcome. Using paired Luminex-based multiplex technique and flow cytometry, we characterized longitudinal dynamics of soluble immune mediators (SIM, n=50) and immune cells in the blood of 244 patients at 8 visits over one year: before, 7/14/21/28 days, 3/6/12 months after pLT. The unsupervised clustering of patients based on SIM profiles revealed 6 unique SIM signatures associated with clinical outcome. From 3 signatures linked to improved outcome, one was associated with one-year-long rejection-free survival and stable graft function and was characterized by low levels of pro-inflammatory (CXCL8/9/10/12, CCL7, SCGF-β, sICAM-1), high levels of regenerative (SCF, TNF-β), and pro-apoptotic (TRAIL) SIM (all, p<0.001, fold change >100). Of note, this SIM signature appeared two weeks after pLT and remained stable over the entire year, pointing towards its potential as a novel early biomarker for minimizing or weaning immunosuppression. In the blood of these patients, a higher frequency of CD56bright NK cells (p<0.01), a known hallmark also associated with operationally tolerant pLT patients, was detected. The concordance of the model for prediction of rejection based on identified SIM signatures was 0.715, and 0.795, in combination with living-related transplantation as co-variate, respectively. SIM blood signatures may enable the non-invasive and early assessment of rejection risks in the first year after pLT, paving the way to improved therapeutic options.

The Prospective Impact of Some Edible Herbal Extracts on Cancer Cell Viability, Biochemical, and Cellular Immune Mediators In vitro

The Prospective Impact of Some Edible Herbal Extracts on Cancer Cell Viability, Biochemical, and Cellular Immune Mediators In vitro

Heterologous DNA Prime- Subunit Protein Boost with Chikungunya Virus E2 Induces Neutralizing Antibodies and Cellular-Mediated Immunity

Chikungunya virus (CHIKV) has become a significant public health concern due to the increasing number of outbreaks worldwide and the associated comorbidities. Despite substantial efforts, there is no specific treatment or licensed vaccine against CHIKV to date. The E2 glycoprotein of CHIKV is a promising vaccine candidate as it is a major target of neutralizing antibodies during infection. In this study, we evaluated the immunogenicity of two DNA vaccines (a non-targeted and a dendritic cell-targeted vaccine) encoding a consensus sequence of E2CHIKV and a recombinant protein (E2*CHIKV). Mice were immunized with different homologous and heterologous DNAprime-E2* protein boost strategies, and the specific humoral and cellular immune responses were accessed. We found that mice immunized with heterologous non-targeted DNA prime- E2*CHIKV protein boost developed high levels of neutralizing antibodies, as well as specific IFN-γ producing cells and polyfunctional CD4+ and CD8+ T cells. We also identified 14 potential epitopes along the E2CHIKV protein. Furthermore, immunization with recombinant E2*CHIKV combined with the adjuvant AS03 presented the highest humoral response with neutralizing capacity. Finally, we show that the heterologous prime-boost strategy with the non-targeted pVAX-E2 DNA vaccine as the prime followed by E2* protein + AS03 boost is a promising combination to elicit a broad humoral and cellular immune response. Together, our data highlights the importance of E2CHIKV for the development of a CHIKV vaccine.

Cellular T-cell immune response profiling by tetravalent dengue subunit vaccine (DSV4) candidate in mice.

While most vaccines aim to develop a solid humoral and neutralizing antibody response against the pathogen, an effective vaccine candidate should be able to stimulate both the B-cell mediated humoral immunity, and T-cell mediated cellular immunity. The focus of vaccinology is rapidly gaining to generate T cell responses, which can mediate pathogen clearance and help B cells leading to protective antibody responses. Here we evaluate the cellular immune response of the pre-clinical tetravalent dengue subunit vaccine candidate, DSV4, in mice. While we have shown previously that DSV4 induces type-specific neutralizing antibody responses in mice, in this study, we show that the vaccine candidate DSV4 well induces dengue-specific T- cell responses evaluated by their ability to produce IFN-γ. In addition to IFN-γ secretion by both CD4+ and CD8+ T-cells in immunized mice, we observed that DSV4 also induces a higher frequency and cytokine functions of follicular CD4+ helper T-cells (TFH). These cytokines lead to an efficient germinal center reaction and potent B cell antibody response. Apart from TFH response, DSV4 stimulated Type 1 T helper cells (TH1) which is characteristic of a viral infection leading to secretion of pro-inflammatory cytokines and phagocyte-dependent protective immune responses. Our study highlights that DSV4 can mediate both arms of adaptive immunity-humoral and cell-mediated immunity in mice. By elucidating vaccine-specific T cell response, our work has implications in showing DSV4 as an effective, type-specific and safe dengue vaccine candidate.

The immunomodulatory role of withania somnifera (L.) dunal in inflammatory diseases.

Withania somnifera (L.) Dunal (Solanaceae) (also known as Ashwagandha) is a botanical drug that has been used for centuries to treat many chronic diseases like high blood pressure, arthritis, diabetes, Alzheimer's disease, and depression. As many botanical drugs, w. Somnifera possesses anti-inflammatory, antioxidant, anticarinogenic, anti-diabetic, and anti-asthmatic properties. W. somnifera is often compared to the ginseng plant due to its ability to reduce stress, improve cognitive functions (e.g., memory), and promote a healthy immune system. It promotes immunomodulatory effects whose function is to balance the humoral and cellular responses of the adaptive immune system. The therapeutic effect of w. Somnifera is attributed to active ingredients like alkaloids, steroidal lactones (such as withanolides, withaferins), and steroidal saponins. Although w. Somnifera is safe and highly recommended for treating various diseases, the current knowledge and understanding of its operational mechanisms are limited. One of the proposed mechanisms states that w. Somnifera promotes cellular-mediated immunity or initiates chemical interactions that contribute to therapeutic effects. Withania somnifera has been shown to play a significant role in immunological diseases by modulating several cytokines, increasing T-cell proliferation and enhancing macrophages functions. In this review, we will discuss the latest therapeutic effects of w. Somnifera on a number of diseases through modulating immunological markers and which specific components of w. Somnifera induce these therapeutic activities. We will also focus on the chemical properties in w. Somnifera components and their immunomodulatory role in type 2 allergic diseases where type 2 inflammation is highly imbalanced.

Recombinant Protein Vaccines Formulated with Enantio-Specific Cationic Lipid R-DOTAP Induce Protective Cellular and Antibody-Mediated Immune Responses in Mice.

Adjuvants are essential components of subunit vaccines added to enhance immune responses to antigens through immunomodulation. Very few adjuvants have been approved for human use by regulatory agencies due to safety concerns. Current subunit vaccine adjuvants approved for human use are very effective in promoting humoral immune responses but are less effective at promoting T-cell immunity. In this study, we evaluated a novel pure enantio-specific cationic lipid 1,2-dioleoyl-3-trimethylammonium-propane (R-DOTAP) as an immunomodulator for subunit vaccines capable of inducing both humoral- and cellular-mediated immunity. Using recombinant protein antigens derived from SARS-CoV2 spike or novel computationally optimized broadly reactive influenza antigen (COBRA) proteins, we demonstrated that R-DOTAP nanoparticles promoted strong cellular- and antibody-mediated immune responses in both monovalent and bivalent vaccines. R-DOTAP-based vaccines induced antigen-specific and polyfunctional CD8+ and CD4+ effector T cells and memory T cells, respectively. Antibody responses induced by R-DOTAP showed a balanced Th1/Th2 type immunity, neutralizing activity and protection of mice from challenge with live SARS-CoV2 or influenza viruses. R-DOTAP also facilitated significant dose sparing of the vaccine antigens. These studies demonstrate that R-DOTAP is an excellent immune stimulator for the production of next-generation subunit vaccines containing multiple recombinant proteins.

Rift Valley Fever vaccine strategies: Enhanced stability of RVF Clone 13

Rift Valley Fever virus (RVFV) causes the zoonotic RVF disease, which results in substantial economic losses in livestock industries. Regular vaccination of livestock against RVF is necessary to generate long-term immunity and avoid the loss of livestock. The live attenuated vaccine based on Clone 13 virus strain has been used to reduce the negative impact of RVF disease. The vaccine strain is heat labile and requires stringent conditions for storage and handling. This research evaluated lactose and sucrose-based stabilizers coupled with lyophilisation to enhance stability of the RVF Clone 13 vaccine strain. The glass transition temperature (Tg) of the sucrose-RVF vaccine was 97.0 °C with average residual moisture of below 2 %. The lactose formulation was characterised with Tg of 83.5 °C and residual moisture of above 2 %. The RVF Clone 13 sucrose-based formulation maintained higher antigen titres during lyophilisation compared to the lactose-formulated vaccine. Cellular-mediated and humoral immunity was evaluated and compared for the two newly formulated vaccines. Pheroid® technology was also investigated as a potential adjuvant and its ability to further enhance the immunogenicity conferred by the RVF Clone 13 vaccine formulations in Merino sheep. No adverse reactions were observed following injection of the vaccine formulations in mice, guinea pigs and Merino sheep. Comparable protective humoral immune responses against RVF were obtained for all animals vaccinated with the lactose and sucrose-based stabilisers with and without the Pheroid® adjuvant. No proliferation of CD8+ and CD4+ T-cells as well as expression of IFN-γ was observed for all animals group vaccinated with Pheroid® only. Specific CD8+ IFN-γ+T–cells were expressed at higher levels compared to the CD4+ IFN-γ+T–cells in the RVF Clone 13 vaccines, suggesting that cellular immunity against RVF is through the Class I antigen presentation pathway.

Dual-Responsive Glycopolymers for Intracellular Codelivery of Antigen and Lipophilic Adjuvants.

Traditional approaches to vaccines use whole organisms to trigger an immune response, but they do not typically generate robust cellular-mediated immunity and have various safety risks. Subunit vaccines composed of proteins and/or peptides represent an attractive and safe alternative to whole organism vaccines, but they are poorly immunogenic. Though there are biological reasons for the poor immunogenicity of proteins and peptides, one other key to their relative lack of immunogenicity could be attributed to the poor pharmacokinetic properties of exogenously delivered proteins and peptides. For instance, peptides often aggregate at the site of injection and are not stable in biological fluids, proteins and peptides are rapidly cleared from circulation, and both have poor cellular internalization and endosomal escape. Herein, we developed a delivery system to address the lack of protein immunogenicity by overcoming delivery barriers as well as codelivering immune-stimulating adjuvants. The glycopolymeric nanoparticles (glycoNPs) are composed of a dual-stimuli-responsive block glycopolymer, poly[2-(diisopropylamino)ethyl methacrylate]-b-poly[(pyridyl disulfide ethyl methacrylate)-co-(methacrylamidoglucopyranose)] (p[DPA-b-(PDSMA-co-MAG)]). This polymer facilitates protein conjugation and cytosolic release, the pH-responsive release of lipophilic adjuvants, and pH-dependent membrane disruption to ensure cytosolic delivery of antigens. We synthesized p[DPA-b-(PDSMA-co-MAG)] by reversible addition-fragmentation chain transfer (RAFT) polymerization, followed by the formation and physicochemical characterization of glycoNPs using the p[DPA-b-(PDSMA-co-MAG)] building blocks. These glycoNPs conjugated the model antigen ovalbumin (OVA) and released OVA in response to elevated glutathione levels. Moreover, the glycoNPs displayed pH-dependent drug release of the model hydrophobic drug Nile Red while also exhibiting pH-responsive endosomolytic behavior as indicated by a red blood cell hemolysis assay. GlycoNPs coloaded with OVA and the toll-like receptor 7/8 (TLR-7/8) agonist Resiquimod (R848) activated DC 2.4 dendritic cells (DCs) significantly more than free OVA and R848 and led to robust antigen presentation of the OVA epitope SIINFEKL on major histocompatibility complex I (MHC-I). In sum, the dual-stimuli-responsive glycopolymer introduced here overcomes major protein and peptide delivery barriers and could vastly improve the immunogenicity of protein-based vaccines.

Persistent Antigen Harbored by Alveolar Macrophages Enhances the Maintenance of Lung-Resident Memory CD8+ T Cells.

Lung tissue-resident memory T cells are crucial mediators of cellular immunity against respiratory viruses; however, their gradual decline hinders the development of T cell-based vaccines against respiratory pathogens. Recently, studies using adenovirus (Ad)-based vaccine vectors have shown that the number of protective lung-resident CD8+ TRMs can be maintained long term. In this article, we show that immunization of mice with a replication-deficient Ad serotype 5 expressing influenza (A/Puerto Rico/8/34) nucleoprotein (AdNP) generates a long-lived lung TRM pool that is transcriptionally indistinct from those generated during a primary influenza infection. In addition, we demonstrate that CD4+ T cells contribute to the long-term maintenance of AdNP-induced CD8+ TRMs. Using a lineage tracing approach, we identify alveolar macrophages as a cell source of persistent NP Ag after immunization with AdNP. Importantly, depletion of alveolar macrophages after AdNP immunization resulted in significantly reduced numbers of NP-specific CD8+ TRMs in the lungs and airways. Combined, our results provide further insight to the mechanisms governing the enhanced longevity of Ag-specific CD8+ lung TRMs observed after immunization with recombinant Ad.

Responses of Humoral and Cellular Immune Mediators in BALB/c Mice to LipX (PE11) as Seed Tuberculosis Vaccine Candidates.

A member of the pe/ppe gene family, lipX (pe11), is capable of directing persistent Mycobacterium tuberculosis and avoiding host immune responses. Some studies have indicated that LipX (PE11) can detect humoral antibodies in tuberculosis patients. Hence, information on immune mediators' responses to this protein is essential to understand its protective efficacy against M. tuberculosis infections. This study aimed to examine the response of immune mediators to pCDNA3.1-lipX expression in vivo. In the experiment, pCDNA3.1-lipX was injected into BALB/c strain male mice aged between 6 and 8 weeks, and they were compared to groups injected with pCDNA3.1 and without injection. The injection was carried out three times intramuscularly every two weeks. Blood was taken retro-orbitally and used for humoral response analysis by Western blotting against LipX-His protein. Simultaneously, the splenocytes were cultured and induced with LipX-His protein for cellular immunity analyses. Our study showed that the recombinant DNA of pCDNA3.1-lipX induced a humoral and cellular immune response, especially in IL-4, IL-12, and IFN-γ, which are the primary cellular responses to M. tuberculosis infections. However, additional studies, such as a challenge study, are needed to strengthen the argument that this plasmid construction is feasible as a tuberculosis seed vaccine candidate.

Effect of propolis supplementation and breed on growth performance, immunity, blood parameters and cecal microbiota in growing rabbits.

ObjectiveThe present study was conducted to investigate the potential effects of dietary supplemented propolis in two growing rabbit breeds on growth performance, immune response, blood parameters, carcass characteristics, and cecal microflora composition.MethodsA total of 90 growing rabbits aged 6 weeks from two breeds (V-line and Jabali) were randomly allocated to 3 dietary propolis experimental treatments. The experimental treatments consisted of a 2×3 factorial arrangement with two rabbit breeds and three levels of dietary propolis supplementation (0, 250 mg/kg, and 500 mg/kg). Each sub-treatment has 15 rabbits. The experimental period lasted six weeks.ResultsThere were no significant differences in growth performance and carcass characteristics due to propolis administration. Propolis supplementation at a high level significantly increased (linear; p<0.05) cellular-mediated immunity compared with the unsupplemented group. Furthermore, the rabbits receiving propolis exhibited a significant increase (linear and quadratic; p<0.03) in IgM immunoglobulins compared to the control. The current study provides further evidence that the dietary inclusion of propolis can significantly reduce pathogenic bacterial colonization in growing rabbits. The total count of microflora, E. coli, and Salmonella spp. was significantly lower (linear; p<0.01) in supplemented rabbit groups compared to the control group according to the microbiological analysis of cecal digesta. Based on breed effect, the results indicated that Jabali rabbits (local) performed better than V-line rabbits (foreign) in the majority of the studied traits.ConclusionDietary propolis is promising for further investigation into improving intestinal health and enhancing immunity in growing rabbits.

Prohemocytes are the main cells infected by dengue virus in Aedes aegypti and Aedes albopictus

BackgroundThe primary disease vectors for dengue virus (DENV) transmission between humans are the mosquitoes Aedes aegypti and Aedes albopictus, with Ae. aegypti population size strongly correlated with DENV outbreaks. When a mosquito is infected with DENV, the virus migrates from the midgut to the salivary glands to complete the transmission cycle. How the virus crosses the hemocoel, resulting in systemic infection, is still unclear however. During viral infection and migration, the innate immune system is activated in defense. As part of cellular-mediated immunity, hemocytes are known to defend against bacteria and Plasmodium infection and may also participate in defending against DENV infection. Hemocytes are categorized into three cell types: prohemocytes, granulocytes, and oenocytoids. Here, we investigated which hemocytes can be infected by DENV and compare hemocyte infection between Ae. aegypti and Ae. albopictus.MethodsHemocytes were collected from Ae. aegypti and Ae. albopictus mosquitoes that were intrathoracically infected with DENV2-GFP. The collected hemocytes were then identified via Giemsa staining and examined microscopically for morphological differences and viral infection.ResultsAll three types of hemocytes were infected by DENV, though the predominantly infected cell type was prohemocytes. In Ae. aegypti, the highest and lowest infection rates at 7 days post infection occurred in prohemocytes and granulocytes, respectively. Prohemocytes were also the primary infection target of DENV in Ae. albopictus, with similar infection rates across the other two hemocyte groups. The ratios of hemocyte composition did not differ significantly between non-infected and infected mosquitoes for either species.ConclusionsIn this study, we showed that prohemocytes were the major type of hemocyte infected by DENV in both Ae. aegypti and Ae. albopictus. The infection rate of prohemocytes in Ae. albopictus was lower than that in Ae. aegypti, which may explain why systemic DENV infection in Ae. albopictus is less efficient than in Ae. aegypti and why Ae. albopictus is less correlated to dengue fever outbreaks. Future work in understanding the mechanisms behind these phenomena may help reduce arbovirus infection prevalence.Graphical

Differential antibody response to COVID-19 vaccines across immunomodulatory therapies for multiple sclerosis

BackgroundPrior studies suggest reduced humoral response to COVID-19 vaccination in immunosuppressed populations. Disease modifying therapies (DMTs) for multiple sclerosis (MS) have variable immunomodulatory effects, and limited data are available for all DMTs. We aimed to determine the impact of DMTs on antibody response to COVID-19 vaccination among MS patients. MethodsPatients with documented COVID-19 vaccination dates and anti-spike antibody results post-vaccination were identified between March-August 2021. Clinical data were retrospectively abstracted from chart review. Deidentified data were analyzed to evaluate antibody response, and multivariable logistic regression analyses were used to identify clinical and demographic predictors of antibody response. Data analysis was completed with SAS Studio, v3.8. ResultsA total of 353 individuals had documented COVID-19 vaccine and antibody test dates (58% Pfizer, 38% Moderna, and 4% Johnson & Johnson). Of these 353 patients, 72% developed antibodies, with a mean antibody test interval of 53 days (median 46) post final vaccine dose. 100% of those on no DMT (n = 34), injectables (n = 20), teriflunomide (n = 10), natalizumab (n = 71), and 97.8% of those on fumarates (n = 46/47) had a positive antibody result. One patient on cladribine (n = 1) had a negative antibody result. Of those on sphingosine-1 phosphate (S1P) modulators, 72.4% (n = 21/29) had a positive antibody result. Of those on anti-CD20 therapies, 37.6% (n = 53/141) had a positive antibody result. Multivariate modeling of the total cohort showed anti-CD20 therapy was significantly associated with lower odds of positive antibody response (OR = 0.024, 95% CI 0.01;0.05, p < 0.0001). Among S1P modulators, increased duration of therapy, and not lymphopenia, may be associated with lower odds of positive antibody response. Multivariate modeling of anti-CD20 therapies showed therapy duration < 1 year (OR 8.14, 95% CI 2.896;22.898 p < .0001) and prior COVID-19 infection (OR = 3.95, 95% CI 1.137;13.726, p = .03) were significantly associated with higher odds of a positive antibody response. In patients with recent B-cell data, mean B-cell count was higher in antibody-positive individuals compared to antibody-negative (32.9 vs. 3.9 cells, p = .0056). ConclusionMS DMTs had variable impact on antibody response with mRNA and viral vector COVID-19 vaccines. All patients on no DMT, interferons, glatiramer acetate, teriflunomide, natalizumab, and nearly all on fumarates had positive antibody responses post-vaccine. S1P modulators and anti-CD20 therapies attenuated antibody response post-vaccine. For patients on anti-CD20 therapies, shorter duration of therapy and prior COVID-19 infection predicted positive antibody response. Further studies are needed to determine clinical significance of antibody testing, development of cellular mediated immunity, and benefits of booster vaccinations.

Separation and phagocytosis analysis of peripheral blood leukocytes from Qihe crucian carp Carassius auratus

The phagocytic function of peripheral blood leukocytes (PBL) in teleost fish is the bridge between cellular-mediated and humoral immunity and plays a central role in the immune system of the teleost fish. However, the phagocytic ability of PBL from Qihe crucian carp Carassius auratus remains unclear. This study aimed to build a convenient separation method to obtain high purity PBL from Qihe crucian carp and to explore their phagocytic characteristics. The results showed that the PBL of Qihe crucian carp (QPBL) could be obtained effectively by 1.085 g/ml percoll separation liquid. Most isolated QPBL were round or elliptical, with horseshoe-shaped, reniform, or irregular nucleus. Finger-like or filamentous protrusions were present on the cell surface, and some cells had a long pseudopod. According to the results of scanning electron microscopy (SEM), transmission electron microscopy (TEM), and fluorescence microscope, fluoresbrite carboxylate YG microspheres (1 μm) could be effectively phagocytosed by QPBL, and various phagocytic stages could be observed. The phagocytic cells in QPBL have morphology similar to those of “professional” phagocytes (monocyte and granulocyte). Notably, the percentage of cells, which can engulf microspheres, increased to approximately 22% after incubation with microspheres for 90 min, and it showed no obvious changes until 180 min. Furthermore, two important phagocytosis regulatory protein genes, Rab5 and Rab7, were seriously up-regulated during the phagocytosis in QPBL. Taken together, an efficient and convenient separation method for isolating QPBL was established, and the isolated QPBL had a high phagocytic activity towards the external particles. The results could provide the basic foundation for further study of the immune function of PBL in Qihe crucian carp and other teleost fish.

Protection against avian coronavirus conferred by oral vaccination with live bacteria secreting LTB-fused viral proteins

The devastating impact of infectious bronchitis (IB) triggered by the IB virus (IBV), on poultry farms is generally curbed by livestock vaccination with live attenuated or inactivated vaccines. Yet, this approach is challenged by continuously emerging variants and by time limitations of vaccine preparation techniques. This work describes the design and evaluation of an anti-IBV vaccine comprised of E. coli expressing and secreting viral spike 1 subunit (S1) and nucleocapsid N-terminus and C-terminus polypeptides fused to heat-labile enterotoxin B (LTB) (LS1, LNN, LNC, respectively). Following chicken oral vaccination, anti-IBV IgY levels and cellular-mediated immunity as well as protection against virulent IBV challenge, were evaluated 14 days following the booster dose. Oral vaccination induced IgY levels that exceeded those measured following vaccination with each component separately. Following exposure to inactivated IBV, splenocytes isolated from chicks orally vaccinated with LNN or LNC -expressing bacteria, showed a higher percentage of CD8+ cells as compared to splenocytes isolated from chicks vaccinated with wild type or LTB-secreting E. coli and to chicks subcutaneously vaccinated. Significant reduction in viral load and percent of shedders in the vaccinated chicks was evident starting 3 days following challenge with 107.5 EID50/ml virulent IBV. Taken together, orally delivered LTB-fused IBV polypeptide-expressing bacteria induced virus-specific IgY antibody production and was associated with significantly shorter viral shedding on challenge with a live IBV. The proposed vaccine design and delivery route promise an effective and rapidly adaptable means of protecting poultry farms from devastating IB outbreaks.

The Profile of Type 1 Leprosy Reaction at Leprosy Division of Dermatology and Venerology Outpatient Clinic of Dr. Soetomo General Academic Hospital, Surabaya, Indonesia

Background: Leprosy is a chronic infectious disease caused by Mycobacterium leprae. Type 1 leprosy reaction is a delayed hypersensitivity reaction caused by the increased response of cellular-mediated immunity to the Mycobacterium leprae antigen on the skin and nerves with a reversal result. The clinical manifestation includes inflammation which can cause skin and nerve lesions, swell, to permanent disabilities. Purpose: To describe the demographic and clinical profile of type 1 leprosy reaction at the Leprosy Division of the Dermatology and Venerology Outpatient Clinic of Dr. Soetomo General Academic Hospital in 2017–2019. Methods: This was a descriptive study. We used secondary data from the medical records of leprosy patients at the Leprosy Division of Dermatology and Venereology outpatient clinic, Dr. Soetomo General Academic Hospital Surabaya, from January 2017 to December 2019. Result: Out of 364 patients, 65 (17.9%) had type 1 reactions. They were mostly in productive age at 35–55 years old (56.9%). The patients were predominantly male (75.4%), with normal nutritional status (98.5%) and negative bacterial index (72.3%). The most common types of leprosy were BB (Borderline) with 61.6% and BL (Borderline Lepromatous) with 20.8%. All patients took WHO (World Health Organization) MDT (Multi Drug Therapy) MB (Multi-Bacillary). Conclusion: The profile of type 1 leprosy reaction at the Leprosy Division of Dermatology and Venerology Outpatient Clinic of Dr. Soetomo General Academic Hospital in 2017–2019 shows an average data as follows: age 35–55 years, male, normal nutritional status, negative bacterial index, leprosy type BB.

Risk Factors of Type 1 Leprosy Reaction in Leprosy Patients attending Leprosy Division of Dermatology and Venereology Outpatient Clinic of Dr Soetomo General Hospital in 2017–2019: A Retrospective Study

BACKGROUND: Type 1 leprosy reaction is a delayed hypersensitivity reaction caused by increased response of cellular-mediated immunity to Mycobacterium leprae. Manifestations include skin and nerve lesions, edema, and permanent disabilities. There are several risk factors that should be recognized to prevent disabilities. AIM: The aim of this study was to analyze the relationship of risk factors to the occurrence of type 1 leprosy reaction in leprosy patients treated at the Outpatient Clinic of Dr. Soetomo General Hospital. METHODS: This study was an analytical study with retrospective observational study design. Data were secondary from the medical records of leprosy patients at the Outpatient Clinic of Dr. Soetomo General Hospital from January 2017 to December 2019. RESULTS: Out of 364 patients in the Outpatient Clinic, 190 (52.2%) had leprosy without a reaction and 65 (17.9%) had type 1 reaction. Analysis showed that age, leprosy type, and treatment regimen were significantly associated with the incidence of type 1 reaction (p = 0.023; 0.003 and 0.004, respectively), with the leprosy type as the most dominant risk factor. Age 15–34 years old; leprosy types BB, BL, and BT; and the MB MDTL therapeutic regimen are risk factors for the occurrence of type I leprosy reaction. CONCLUSION: There is a statistically significant correlation between the risk factor and the occurrence of type 1 leprosy reaction in leprosy patient. The risk factor that has significant correlation is age 15–34 years; leprosy types BB, BL, and BT; and the MB MDTL therapeutic regimen. The most significant risk factor for the occurrence of type 1 leprosy reaction from our study is the type of leprosy (BB, BL, and BT).

B-Cell Lymphoma Patients Have a 10% Risk of Shingles, 7% in Cutaneous T-Cell Lymphoma

Background: Patients with lymphoma can be immunosuppressed from the disease, treatment or both. One indicator of cellular mediated immunity is reactivation of Varicella Zoster (VZV), a painful viral infection estimated to occur to 30% of U.S. population during a lifetime and associated with a particularly morbid post-herpetic neuralgia. Bortezomib and dexamethasone have been shown to be correlated with increased rates of varicella zoster and other viral infections (Kim SJ et al, Clinical lymphoma & myeloma, 2008). Autologous stem cell transplant depletes cellular- and humoral-immunity (Kamber C et al, BMT, 2015). While the main risk factor for VZV reactivation is age (Harpaz R, MMWR Recomm Rep, 2008), the underlying malignancy might play a large role in the risk of VZV reactivation. There is little data comparing the proportion of VZV in multiple myeloma (MM) with other indolent B and T-cell lymphomas. The purpose of this study is to compare varicella infection in MM patients with chronic lymphocytic leukemia (CLL), cutaneous T-cell lymphoma (CTCL), and follicular lymphoma (FL). Given that VZV is a relatively common infection in patients with MM, we hypothesized that varicella reactivation in MM patients occurs at a higher rate than in patients with CLL, CTCL, or FL.Methods: A retrospective chart review of 3,765 patients was conducted. Patients were eligible if they had been diagnosed with 1 of 4 malignancies - MM (1,210), CLL (2,520), CTCL (434), or FL (809). Patients diagnosed with other malignancies i.e. mantle cell lymphoma were excluded, as were patients with multiple malignancies i.e. MM and FL. All patients had been seen at the Ohio State University Wexner Medical Center during their treatment. For each patient, information on age, gender, length of survival, age at diagnosis of hematologic malignancy, and proportion of patients with shingles was collected. Only patients who were diagnosed with shingles after hematologic cancer diagnosis were included as positive. Associations between malignancy type and development of shingles were analyzed by a chi-square test, and logistic regression was used to calculate odds ratios for shingles between MM and the other malignancies, controlling for age as a possible confounder. Median survival time was calculated in years using the Kaplan-Meier method, calculated as the time from cancer diagnosis to date of last contact if alive, or date of death if deceased. Seven patients were excluded from survival analysis due to having the same date of diagnosis as date of last contact. This study was approved by the cancer institutional review board.Results: Patient demographic information is found in Table 1. Of the 1,210 MM patients, 127 developed shingles after diagnosis (10.5%); of 2520 CLL patients, 326 were positive (12.9%); of 434 CTCL patients, 33 were positive (7.6%); of 809 FL patients, 78 were positive (9.6%). At a significance level of a=0.05, there was a significant association between cancer type and shingles (χ2=15.59, df=3, p-value=0.0014). Odds ratios comparing MM to each of the other lymphomas were not found to be significant. The median survival time in years was 8.8 for MM, 20.7 for CLL, 25.1 for CTCL, and 24.6 for FL (Figure 1).Conclusion: Patients with MM had the lowest median survival (8.8 years), and a significant association between cancer type and shingle incidence was found, with CTCL patients having the lowest incidence of shingles (7.6%) within the retrospective period. However, the odds ratios comparing MM to the other 3 lymphomas was not significant, showing that the MM patients did not have significantly higher rates of shingles as originally hypothesized. These results demonstrate the differences in viral infection incidence and disease course between hematologic malignancies, and that antiviral strategies more specialized to T and B cell lymphomas may be important in reducing the risk of viral infection. [Display omitted] DisclosuresHofmeister:Celgene: Research Funding; Adaptive Biotechnologies: Honoraria, Membership on an entity's Board of Directors or advisory committees; Karyopharm: Research Funding; Takeda: Research Funding; Thrassos: Honoraria, Membership on an entity's Board of Directors or advisory committees; Janssen: Research Funding; Bristol-Myers Squibb: Research Funding; Roche: Research Funding.

Exploring Onchocerca volvulus Cysteine Protease Inhibitor for Multi-epitope Subunit Vaccine Against Onchocerciasis: An Immunoinformatics Approach

Onchocerciasis, caused by Onchocerca volvulus, affects more than 37 million people worldwide. Despite the progress achieved with mass drug distribution, suitable vaccines against onchocerciasis are needed to effectively eliminate the infection. The O. volvulus cysteine protease inhibitor (onchocystatin) is an immuno-dominant antigen detected in O. volvulus infections, capable of inducing protective immunity. Here, we explore the onchocystatin for a multi-epitope subunit vaccine candidate targeted against onchocerciasis. A multi-epitope vaccine candidate composed of RS-09 as adjuvant, a CD8+ T cell peptide, a CD4+ T cell peptide and a B cell peptide concatenated with suitable linkers was computationally constructed. Immune simulation of the vaccine response predicted several aspects of antibody-dependent and cellular-mediated immunity with accompanied B cell and helper T cell immune memory development. The levels of lFN-γ and IL-2 were also predicted to be elevated. Collectively, our results suggest that the multi-epitope vaccine construct has the potential to mimic the natural immunity targeted against onchocerciasis and other related filarial infections, and should be considered for further experimental validations.