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Crimean-Congo hemorrhagic fever (CCHF) is a zoonotic disease caused by Crimean-Congo hemorrhagic fever virus (CCHFV), which can cause severe clinical disease and even death in humans. In recent years, the disease has spread to a wider area, posing a major public health threat to China as well as the Middle East, Europe and Africa, and there is no safe and effective vaccine to prevent the disease. Recently, it has been shown that using the Zera fusion to target proteins can enhance immunogenicity and improve the potential for developing viral vaccines. Based on this finding, in this study, two vaccine candidates, Zera-Gn and Zera-Np, were prepared using an insect baculovirus system expressing CCHFV glycoprotein (Gn) and nucleocapsid protein (Np) fused with Zera tags, and evaluated for immunogenicity in BALB/c mice. The obtainedresults showed that both Zera-Gn and Zera-Np recombinant nanoparticles were successfully expressed, and Zera-Gn had good induction of humoral and cellular immunity in mice, and its immunogenicity was significantly higher than that of Zera-Np. The results indicated that Zera-Gn self-assembled nanoparticles prepared by fusing Zera tags with CCHFV spike-in protein Gn have the potential to be a candidate vaccine for CCHF, and this study provides a reference for the development of Zera self-assembled nanoparticle vaccine for CCHF.

Crimean-Congo hemorrhagic fever (CCHF), which has a fatality rate of 20–30%, is widely prevalent in several regions in Asia, Europe, and Africa and has spread to a wider range of areas in recent years. At present, there is a lack of safe and effective vaccines for the prevention of CCHF. In this study, we prepared three vaccine candidates, rvAc-Gn, rvAc-Np, and rvAc-Gn-Np, that encoded the CCHF virus (CCHFV) glycoprotein Gn and the nucleocapsid protein (Np) on the surface of baculovirus using an insect baculovirus vector expression system (BVES) and evaluated their immunogenicity in BALB/c mice. The experimental results showed that both CCHFV Gn and Np were expressed by the respective recombinant baculoviruses and anchored to the viral envelope. BALB/c mice were immunized, and all three recombinant baculoviruses showed significant humoral immunity. At the cellular level, the level of immunity in the rvAc-Gn group was significantly higher than that in the rvAc-Np and rvAc-Gn-Np groups, and the rvAc-Gn-Np coexpression group exhibited the lowest level of cellular immunity. In conclusion, the strategy of coexpressing Gn and Np in the baculovirus surface display system did not result in improvements in immunogenicity, whereas the recombinant baculovirus displaying Gn alone could induce significant humoral and cellular immunity in mice, indicating that rvAc-Gn has potential as a CCHF vaccine candidate. This study thus provides new ideas for the development of a CCHF baculovirus vaccine. [ABSTRACT FROM AUTHOR]

Comprehensive Summary: Proteus species especially Proteus mirabilis and Proteus vulgaris are zoonotic pathogens which can cause public health disease. Owing to their antibiotic‐resistance, developing vaccines against these pathogens is urgently required. Herein, we describe the first synthesis of the common O‐antigen of Proteus mirabilis OE and Proteus vulgaris TG 103. The repeating unit incorporates two challenging 1,2‐cis‐glycosidic bonds: the 1,2‐cis‐2‐acetamido‐2‐deoxy‐glucosidic bonds were successfully constructed by use of conformation‐restrained 2‐nitroglucal donor under bifunctional organothiourea catalysis; while the 1,2‐cis‐2‐acetamido‐2‐deoxy‐galactosidic bonds were formed by use of di‐tert‐butylsilylidene protected 2‐azidogalactose donors. The synthetic fragments were screened by glycan microarray with immunized rabbit sera against purified LPS from Proteus mirabilis O54. The results revealed that the synthetic common O‐antigens both hexasaccharide 2 and nonasaccharide 3 can strongly bind with the IgG antibodies (Abs) in the sera, which is highly valuable for further immunological investigation in synthetic carbohydrate‐based vaccine development. [ABSTRACT FROM AUTHOR]

Tuberculosis (TB) remains a serious global health problem. Despite the widespread use of the Mycobacterium bovis bacillus Calmette-Guerin (BCG) vaccine, the primary factor for the TB pandemic and deaths is adult TB, which mainly result from endogenous reactivation of latent Mycobacterium tuberculosis (MTB) infection. Improved new TB vaccines with eligible safety and long-lasting protective efficacy remains a crucial step toward the prevention and control of TB. In this study, five immunodominant antigens, including three early secreted antigens and two latency associated antigens, were used to construct a single recombinant fusion protein (Epera013f) and a protein mixture (Epera013m). When formulated with aluminum adjuvant, the two subunit vaccines Epera013m and Epera013f were administered to BALB/c mice. The humoral immune responses, cellular responses and MTB growth inhibiting capacity elicited after Epera013m and Epera013f immunization were analyzed. In the present study, we demonstrated that both the Epera013f and Epera013m were capable of inducing a considerable immune response and protective efficacy against H37Rv infection compared with BCG groups. In addition, Epera013f generated a more comprehensive and balanced immune status, including Th1, Th2 and innate immune response, over Epera013f and BCG. The multistage antigen complex Epera013f possesses considerable immunogenicity and protective efficacy against MTB infection ex vivo indicating its potential and promising applications in further TB vaccine development. [ABSTRACT FROM AUTHOR]

Astragaloside VII (AST VII), a triterpenic saponin isolated from Astragalus species, shows promise as a vaccine adjuvant, as it supported a balanced Th1/Th2 immune response in previous in vivo studies. However, the underlying mechanisms of its adjuvant activity have not been defined. Here, we investigated the impact of AST VII and its newly synthesized semi-synthetic analogs on human whole blood cells, as well as on mouse bone marrow-derived dendritic cells (BMDCs). Cells were stimulated with AST VII and its derivatives in the presence or absence of LPS or PMA/ionomycin and the secretion of cytokines and the expression of activation markers were analyzed using ELISA and flow cytometry, respectively. AST VII and its analogs increased the production of IL-1β in PMA/ionomycin-stimulated human whole blood cells. In LPS-treated mouse BMDCs, AST VII increased the production of IL-1β and IL-12, and the expression of MHC II, CD86, and CD80. In mixed leukocyte reaction, AST VII and derivatives increased the expression of the activation marker CD44 on mouse CD4+ and CD8+ T cells. In conclusion, AST VII and its derivatives strengthen pro-inflammatory responses and support dendritic cell maturation and T cell activation in vitro. Our results provide insights into the mechanisms of the adjuvant activities of AST VII and its analogs, which will be instrumental to improve their utility as a vaccine adjuvant. [ABSTRACT FROM AUTHOR]

On the territory of Russia, stem rust is found annually in the main grain-producing regions: North Caucasus, Western Siberia, the Volga region, the Central Chernozem region, the Trans-Urals, and Moscow oblast. Therefore, annual measures to monitor the spread and forecast of stem rust are especially relevant to prevent possible epiphytoties. The object of research was 25 released winter wheat varieties obtained at Lukyanenko National Grain Center (Krasnodar), two winter wheat varieties obtained at the Federal Rostov Agrarian Scientific Center (Rostov oblast), and one winter wheat variety obtained at the Donskoy Agrarian Scientific Center (Rostov oblast). The evaluation was carried out at a field station with artificial infection by stem rust of the Federal Research Center of Biological Plant Protection (Krasnodar) in 2018–2020. The evaluation criteria were the type of plant reaction to infection expressed in points, the degree of damage expressed in percent, and the area under the disease progress curve expressed in conventional units. Plant inoculation, the determination of the type of reaction, and the degree of damage were carried out according to standard methods. According to the results of the immunological evaluation, the following varieties showed resistance characterized by the degree of damage up to 10% and the reaction type was one point: Antonina, Afina, Bezostaya 100, Vassa, Gratsiya, Zhiva, Kuren, and Utrish. Nine varieties (Adel, Dolya, Dmitry, Eremeevna, Sila, Trio, Yubileinaya 100, Yuka, Etnos) were moderately resistant, having been affected up to 30%, and the reaction type was two points. Seven varieties (Grom, Gubernator Dona, Don mira, Esaul, Kalym, Stanichnaya, Tanya) were classified as moderately susceptible, the degree of damage was up to 60%, and the reaction type was three points. Three varieties (Ermak, Laureat, Lebed) were susceptible, they were affected above 60%, and the reaction type was four points. Varieties that showed resistance to Puccinia graminis under conditions of the field station, revealed by artificial infection of stem rust, can be recommended for the use in agricultural production in regions with constant manifestation of the pathogen and as sources of resistance for inclusion in breeding programs. [ABSTRACT FROM AUTHOR]

The objective of this study was to develop ritonavir (RTV) nanosuspensions (NSs) by microfluidization method. Particle size (PS) measurements were performed by photon correlation spectroscopy. Amorphous properties of the particles were evaluated by X-ray diffraction (XRD) and scanning electron microscopy (SEM). The dissolution studies were conducted in fed state simulated intestinal fluid (FeSSIF) medium. The flow cytometry was utilized to determine the lymphocyte sub-groups and immune response of NSs. RTV NSs were obtained with 400–500 nm PS. The crystal properties of RTV remain unchanged. The solubility of NS was enhanced five times. 57% and 18% of RTV were dissolved in FeSSIF medium for NSs and coarse powder. According to immunological studies, the prepared NSs did not significantly alter the ratio of CD4+/CD8+. Therefore, NSs may be a beneficial approach for the oral administration of RTV. [ABSTRACT FROM AUTHOR]

Vaccine adjuvants are key for optimal vaccine efficacy, increasing the immunogenicity of the antigen and potentiating the immune response. Saponin adjuvants such as the carbohydrate-based QS-21 natural product are among the most promising candidates in vaccine formulations, but suffer from inherent drawbacks that have hampered their use and approval as stand-alone adjuvants. Despite the recent development of synthetic derivatives with improved properties, their full potential has not yet been reached, allowing the prospect of discovering further optimized saponin variants with higher potency. Herein, we have designed, chemically synthesized, and immunologically evaluated novel oxime-derivatized saponin adjuvants with targeted structural modifications at key triterpene functionalities. The resulting analogues have revealed important findings into saponin structure-activity relationships, including adjuvant mechanistic insights, and have shown superior adjuvant activity in terms of significantly increased antibody response augmentation compared to our previous saponin leads. These newly identified saponin oximes emerge as highly promising synthetic adjuvants for further preclinical development towards potential next generation immunotherapeutics for future vaccine applications. [ABSTRACT FROM AUTHOR]

Vaccine adjuvants are key for optimal vaccine efficacy, increasing the immunogenicity of the antigen and potentiating the immune response. Saponin adjuvants such as the carbohydrate-based QS-21 natural product are among the most promising candidates in vaccine formulations, but suffer from inherent drawbacks that have hampered their use and approval as stand-alone adjuvants. Despite the recent development of synthetic derivatives with improved properties, their full potential has not yet been reached, allowing the prospect of discovering further optimized saponin variants with higher potency. Herein, we have designed, chemically synthesized, and immunologically evaluated novel oxime-derivatized saponin adjuvants with targeted structural modifications at key triterpene functionalities. The resulting analogues have revealed important findings into saponin structure-activity relationships, including adjuvant mechanistic insights, and have shown superior adjuvant activity in terms of significantly increased antibody response augmentation compared to our previous saponin leads. These newly identified saponin oximes emerge as highly promising synthetic adjuvants for further preclinical development towards potential next generation immunotherapeutics for future vaccine applications.

The article presents a case study of an infant with Kasabach–Merritt phenomenon (KMP), a rare disease with vascular tumors, treated with everolimus (EVL). Topics discussed include the efficacy of EVL in KMP, the risk of pneumocystis pneumonia as a rare but potentially fatal complication of mTOR inhibitors, and the need for immunological evaluations and prophylactic measures during treatment.

Tuberculosis (TB) remains a serious global health problem. Despite the widespread use of the Mycobacterium bovis bacillus Calmette-Guerin (BCG) vaccine, the primary factor for the TB pandemic and deaths is adult TB, which mainly result from endogenous reactivation of latent Mycobacterium tuberculosis (MTB) infection. Improved new TB vaccines with eligible safety and long-lasting protective efficacy remains a crucial step toward the prevention and control of TB. In this study, five immunodominant antigens, including three early secreted antigens and two latency associated antigens, were used to construct a single recombinant fusion protein (Epera013f) and a protein mixture (Epera013m). When formulated with aluminum adjuvant, the two subunit vaccines Epera013m and Epera013f were administered to BALB/c mice. The humoral immune responses, cellular responses and MTB growth inhibiting capacity elicited after Epera013m and Epera013f immunization were analyzed. In the present study, we demonstrated that both the Epera013f and Epera013m were capable of inducing a considerable immune response and protective efficacy against H37Rv infection compared with BCG groups. In addition, Epera013f generated a more comprehensive and balanced immune status, including Th1, Th2 and innate immune response, over Epera013f and BCG. The multistage antigen complex Epera013f possesses considerable immunogenicity and protective efficacy against MTB infection ex vivo indicating its potential and promising applications in further TB vaccine development.

Astragaloside VII (AST VII), a triterpenic saponin isolated from Astragalus species, shows promise as a vaccine adjuvant, as it supported a balanced Th1/Th2 immune response in previous in vivo studies. However, the underlying mechanisms of its adjuvant activity have not been defined. Here, we investigated the impact of AST VII and its newly synthesized semi-synthetic analogs on human whole blood cells, as well as on mouse bone marrow-derived dendritic cells (BMDCs). Cells were stimulated with AST VII and its derivatives in the presence or absence of LPS or PMA/ionomycin and the secretion of cytokines and the expression of activation markers were analyzed using ELISA and flow cytometry, respectively. AST VII and its analogs increased the production of IL-1β in PMA/ionomycin-stimulated human whole blood cells. In LPS-treated mouse BMDCs, AST VII increased the production of IL-1β and IL-12, and the expression of MHC II, CD86, and CD80. In mixed leukocyte reaction, AST VII and derivatives increased the expression of the activation marker CD44 on mouse CD4+ and CD8+ T cells. In conclusion, AST VII and its derivatives strengthen pro-inflammatory responses and support dendritic cell maturation and T cell activation in vitro. Our results provide insights into the mechanisms of the adjuvant activities of AST VII and its analogs, which will be instrumental to improve their utility as a vaccine adjuvant.

Winter barley varieties were studied at different stages of their development to reveal those resistant to the North Caucasian populations of dangerous plant pathogenic fungi and promising for the further use in breeding and grain production. The immunological study included 19 winter barley varieties cultivated in Southern Russia and assessed for resistance to Puccinia hordei, Pyrenophora teres, and Cochliobolus sativus. The study was conducted in Krasnodar during vegetation seasons of 2018/2019 and 2019/2020 under laboratory (climatic chamber, seedling stage) and field (adult plants) conditions using common phytopathological methods. The varieties were ranked according to the level and type of their disease resistance to reveal those with the maximum group resistance. Varieties resistant (R) to barley leaf rust at the seedling stage and characterized by a formation of chlorotic lesions instead of pustules included Karrera and Kubagro-1. Varieties resistant to the net blotch (two-score disease progression level) included Gordey, Pattern, and Sarmat. Varieties resistant to the spot blotch (disease progression between 0.6 and 3.8 scores) included Karrera, Lazar, Madar, and Timofey. Varieties resistant (R) to the spot blotch at the adult plant stage included Gordey, Sprinter, and Timofey (disease progression level did not exceed 10–15%). Varieties moderately resistant to the barley leaf rust (MR, disease progression level between 15 and 30%) included Gordey, Iosif, Karrera, Lais, Pattern, and Sprinter; a moderate resistance to the net blotch (MR, disease progression between 15 and 30%) was observed in vars. Sarmat and Timofey. The group resistance (R and MR) at the seedling stage to P. hordei, P. teres, and C. sativus was observed in vars. Karrera and Sprinter. No plants possessing group resistance to these pathogens at the adult plant stage were revealed. [ABSTRACT FROM AUTHOR]

Recent changes in the epidemiology of meningococcal have been reported and meningococcal group W (MenW) has become the third most prevalent group isolated in Brazil in the last 10 years. In this study we have developed a conjugate vaccine for MenW using a modified reductive amination conjugation method through a covalent linkage between periodate-oxidized MenW non-O-acetylated polysaccharide and hydrazide-activated monomeric tetanus toxoid. Process control of bulks was done by physicochemical analysis including polysaccharide and protein quantification, high performance liquid chromatography – size exclusion chromatography, capillary electrophoresis, and hydrogen nuclear magnetic resonance. Conjugate bulks were best produced with concentration of polysaccharide twice as high as protein, at room temperature, and pH approximately 6.0. A scaled-up bulk (100 mg scale) was formulated and inoculated intramuscularly in mice in a dose–response study (0.1, 0.5, 1.0 and 10.0 µg of polysaccharide/dose). The immunogenicity of conjugate bulks was determined by serum bactericidal assay and ELISA assays of serum from immunized mice. ELISA and SBA titers revealed high titers of IgG and demonstrated the functionality of the antibodies produced in all doses studied 15 days after the third dose. However, significant differences were observed among them by ELISA. In conclusion, this study established the best conditions to produce MenW conjugate bulks and showed the efficacy of the obtained conjugate bulk in induce a good immune response in mice. Further experiments will need to be done to scale up the conjugation reaction and then allow the use of this conjugate in clinical trials. [ABSTRACT FROM AUTHOR]

Objectives: Inborn Errors of Immunity are characterized by infectious conditions and manifestations of immune dysregulation. The diversity of clinical phenotypes can make it difficult to direct the laboratory investigation. This article aims to update the investigation of immunological competence in the context of primary defects of the immune system. Source of data: Searches were carried out on Pubmed to review articles published in the last five years, in English, French or Spanish, using the terms “diagnosis” OR “investigation” AND “immunodeficiency” or “primary immunodeficiency” or “inborn errors of immunity” NOT “HIV”. Recent textbook editions have also been consulted. Summary of findings: The immune system competence investigation should be started based on clinical phenotypes. Relevant data are: characterization of infectious conditions (location, recurrence, types of infectious agents, response to treatment), age during symptom onset and associated manifestations (growth impairment, allergy, autoimmunity, malignancies, fever and signs of inflammation without the identification of infection or autoimmunity) and family history. These data contribute to the selection of tests to be performed. Conclusions: The diagnostic investigation of Inborn Errors of Immunity should be guided by the clinical characterization of patients, aiming to optimize the use of complementary tests. Many diagnoses are attained only through genetic tests, which are not always available. However, the absence of a diagnosis of certainty should never delay the implementation of therapeutic measures that preserve patient life and health.

Objectives: Inborn Errors of Immunity are characterized by infectious conditions and manifestations of immune dysregulation. The diversity of clinical phenotypes can make it difficult to direct the laboratory investigation. This article aims to update the investigation of immunological competence in the context of primary defects of the immune system. Source of data: Searches were carried out on Pubmed to review articles published in the last five years, in English, French or Spanish, using the terms "diagnosis" OR "investigation" AND "immunodeficiency" or "primary immunodeficiency" or "inborn errors of immunity" NOT "HIV". Recent textbook editions have also been consulted. Summary of findings: The immune system competence investigation should be started based on clinical phenotypes. Relevant data are: characterization of infectious conditions (location, recurrence, types of infectious agents, response to treatment), age during symptom onset and associated manifestations (growth impairment, allergy, autoimmunity, malignancies, fever and signs of inflammation without the identification of infection or autoimmunity) and family history. These data contribute to the selection of tests to be performed. Conclusions: The diagnostic investigation of Inborn Errors of Immunity should be guided by the clinical characterization of patients, aiming to optimize the use of complementary tests. Many diagnoses are attained only through genetic tests, which are not always available. However, the absence of a diagnosis of certainty should never delay the implementation of therapeutic measures that preserve patient life and health. [ABSTRACT FROM AUTHOR]

BackgroundGlyco-lipopeptides, a form of lipid-tailed glyco-peptide, are currently under intense investigation as B- and T-cell based vaccine immunotherapy for many cancers. However, the cellular and molecular mechanisms of glyco-lipopeptides (GLPs) immunogenicity and the position of the lipid moiety on immunogenicity and protective efficacy of GLPs remain to be determined.Methods/Principal FindingsWe have constructed two structural analogues of HER-2 glyco-lipopeptide (HER-GLP) by synthesizing a chimeric peptide made of one universal CD4+ epitope (PADRE) and one HER-2 CD8+ T-cell epitope (HER420–429). The C-terminal end of the resulting CD4–CD8 chimeric peptide was coupled to a tumor carbohydrate B-cell epitope, based on a regioselectively addressable functionalized templates (RAFT), made of four α-GalNAc molecules. The resulting HER glyco-peptide (HER-GP) was then linked to a palmitic acid moiety, attached either at the N-terminal end (linear HER-GLP-1) or in the middle between the CD4+ and CD8+ T cell epitopes (branched HER-GLP-2). We have investigated the uptake, processing and cross-presentation pathways of the two HER-GLP vaccine constructs, and assessed whether the position of linkage of the lipid moiety would affect the B- and T-cell immunogenicity and protective efficacy. Immunization of mice revealed that the linear HER-GLP-1 induced a stronger and longer lasting HER420–429-specific IFN-γ producing CD8+ T cell response, while the branched HER-GLP-2 induced a stronger tumor-specific IgG response. The linear HER-GLP-1 was taken up easily by dendritic cells (DCs), induced stronger DCs maturation and produced a potent TLR- 2-dependent T-cell activation. The linear and branched HER-GLP molecules appeared to follow two different cross-presentation pathways. While regression of established tumors was induced by both linear HER-GLP-1 and branched HER-GLP-2, the inhibition of tumor growth was significantly higher in HER-GLP-1 immunized mice (p

The most common and harmful disease of the agricultural crop rice is a “burn” caused by the fungus Magnoporthe grisea (Hebert) Barr, the causative agent of rice blast. The important direction of modern domestic rice breeding is the development of high-yielding varieties resistant to blast. To solve the problem, it is important to search for sources and donors of resistance to the Krasnodar population of the pathogen among ecotypes of different ecological and geographical origin. Evaluation of the rice collection diversity for resistance to blast was carried out both on a natural background and on an infectious-provocative one. Immunological evaluation and phenotyping were carried out in 2015–2017 on 154 varieties of the Oryza sativa L. species from 7 ecological and geographical cultivation zones. Over the years of research, the range of variation in the intensity of the disease development in varieties was in the range from 1.1 to 77.8 %. The differences in the resistance of rice varieties to the pathogen between ecological groups and countries have been found. Most of the studied samples have shown medium resistance, there were isolated 51 resistant forms. Most often stable forms were found among the germplasm from China, Italy, the Philippines and Korea, and the unstable ones were from African countries, Japan, Primorye and Vietnam. Introduced samples resistant to the disease were identified and adapted to soil and climatic conditions and rice cultivation technologies of the Kuban, they were included in the breeding scheme for developing pathogen-resistant rice varieties with the extension of their genetic basis. The article presents data on the variation of morphological traits and the rate of development of plants of international varieties from 24 countries in the conditions of the south of Russia. The results of the comparison of germplasm of domestic and foreign varieties according to the degree of resistance to the pathogen in conditions of natural infection in the field experiment for five years are presented. As a result of the evaluation of plant resistance to the Krasnodar population of the pathogen, the effective genes for resistance to the pathogen for breeding programs of the south of Russia and the molecular genetic analysis of the rice collection variety were determined: Pi-1, Pi-z, Pi-ta, Pi-z5, Pi-9, Pi-5(t), Pi-t, Pi-19.

Cephalopina titillator (C. titillator) is a common worldwide nasal bot fly larval infestation of camels, which belongs to the family Oestridae. This study aimed to evaluate two new immunologic diagnostic techniques; indirect-ELISA and Dot-ELISA, for the screening of C. titillator infestation in camels. Thirty slaughtered camel heads were examined carefully for the presence of C. titillator larvae. One hundred, third-stage larvae (L3), were dissected for the collection of their salivary glands, for the preparation of the salivary gland antigen. Blood samples were obtained for hematological and serological examinations. Results revealed a true prevalence of C. titillator in the sampled camels being 80% (24/30). Infested camels showed a significant reduction in leukocytes (P < 0.0001) and neutrophils (P = 0.045), and a significant increase in eosinophils and monocytes (P < 0.0001). The serological examination estimated apparent prevalence as 80% (24/30) and 90% (27/30) by Dot-ELISA and indirect-ELISA, respectively. Dot-ELISA revealed 100% sensitivity, specificity, and accuracy. While, indirect-ELISA displayed 100% sensitivity, 50% specificity, and 90% accuracy. Dot-ELISA exhibited perfect agreement with the gold standard test, so it could be considered an ideal, simple, and accurate immunologic screening technique for the detection of C. titillator in camels. [ABSTRACT FROM AUTHOR]

We aimed to compare the performance of acellular nerves prepared by different decellularization methods, screening out the optimal decellularization protocol, repairing the sciatic nerve defects in rats by the allogeneic transplantation, and evaluating the effect of regenerative nerve on the function reconstruction. The Sondell, SB‐SDS, TnBP, and the high/low permeation methods were used to decellularize donor nerves. Nerves without any treatment were as the control group. The histological results were evaluated by HE staining and toluidine blue (TB) staining. The proliferation activity of L929 cells was detected by CCK‐8 assay. The adhesion of Schwann cells was observed and quantified by SEM. Balb/c mice were used to evaluate the cellular and humoral immunogenicity of the nerve scaffolds. The rat sciatic nerve defect model was applied to observe the repair effect of acellular nerve scaffold in vivo. To SB‐SDS group, it remained the original state of the nerves, with no observed nucleus and axons, the neurotoxicity grade detected by CCK‐8 being almost 0, and it kept the largest number of Schwann cells adhered to the acellular nerve and the better morphology. Further, it showed that the selected SB‐SDS rats acellular nerve scaffold could promote the nerve repair of the rats by HE staining and TB staining. We could conclude that the acellular nerve matrix prepared by the SB‐SDS method effectively removes the cellular components in the nerve tissue and retains the main components of the extracellular matrix of the nerve tissue, whose rats decellularized nerve scaffold could promote the sciatic nerve repair better. [ABSTRACT FROM AUTHOR]

O-Antigens and core oligosaccharides from bacterial lipopolysaccharides (LPS) are often structurally unique and immunologically active, have become attractive targets in the development of antibacterial vaccines. Structurally well-defined and pure oligosaccharides can be used in identifying protective epitopes of the carbohydrate antigens, which is important for the design of an effective vaccine. Here, the recent progress on chemical synthesis and immunological evaluation of glycans related to O-antigens and core oligosaccharides from bacterial LPS are summarized. [ABSTRACT FROM AUTHOR]

Purpose: To compare plasma concentrations of all lectin pathway (LP) pattern recognition molecules (PRMs) in patients referred for laboratory evaluation due to recurrent infections with healthy individuals. Methods: Patients were divided into categories according to referral: recurrent airway infections (RAI), recurrent abscesses, common variable immunodeficiency (CVID), lung transplantation candidates (LTX), and 'other causes'. LP PRMs (mannose-binding lectin (MBL), collectin liver 1 (CL-L1), H-ficolin, L-ficolin, M-ficolin) and C-reactive protein (CRP) were determined in 332 patients and 150 healthy blood donors using time-resolved immunofluorometric assays. Results: None of the LP PRMs was found in lower concentration in the patient categories; however, several PRMs were detected in higher concentrations. M-ficolin was found in higher concentrations in all patient categories. Patients suffering from RAI had higher concentrations of CL-L1 and H-ficolin. Patients suffering from abscesses exhibited higher concentrations of MBL and CL-L1, whereas LTX had higher concentrations of MBL. Patients with other causes of referral had higher concentrations of MBL and CL-L1. Prevalence of combined deficiencies of PRMs in patient categories and controls did not differ. CRP was used as a marker of ongoing inflammation and was significantly higher among all patient categories. Furthermore, CRP was found to correlate with both M-ficolin and L-ficolin. Conclusion: The results suggest that neither single nor combined deficiencies of LP PRMs are more frequent among patients referred for an immunological evaluation than in healthy individuals. Future studies are needed and should focus on deficiencies of LP PRMs combined with deficiencies in other parts of the immune system. [ABSTRACT FROM AUTHOR]

Aim: The present study evaluated the structural changes resulting from the interaction between a recombinant influenza A virus M2 protein and aluminum hydroxide adjuvant to investigate the antigen for further immunological studies. Materials & methods: Membrane protein II was produced from the H1N1 subtype of human influenza A virus. The interaction between M2 protein and alum inum hydroxide adjuvant was evaluated by physicochemical techniques including scanning electron microscope, UV-Vis spectra, Fourier-transform infrared spectroscopy and circular dichroism spectroscopy. Results: Physicochemical methods showed high-level protein adsorption and accessibility to the effective parts of the protein. Conclusion: It was concluded that M2 protein secondary structural perturbations, including the α-helix-to-β-sheet transition, enhanced its mechanical properties toward adsorption. [ABSTRACT FROM AUTHOR]

Acinetobacter baumannii is considered as a major cause of nosocomial infection worldwide. Various vaccine formulations have been mostly studied based on secreted or surface-exposed proteins of A. baumannii in murine models. Serum resistance proteins are critical virulence factors in bloodstream infections. AbOmpA and PKF are two major factors involved in serum resistance and could be considered as promising vaccine targets. In this study IgG1, IgG2c, Total-IgG concentrations, survival rates and spleen bacterial loads were studied in C57/BL mice model according to PKF, AbOmpA and AbOmpA + PKF vaccine formulations. The findings showed significant raises of IgG2c and Total-IgG in all three vaccinated groups in comparison with the control group. Whereas, there were low concentrations of IgG1 in all immunization plans. Colony counts of mice spleen showed the bacterial load of PKF plan had the most decrease of bacterial load (DBL = 5 log 10 CFU/g). Taken together, this evaluation indicated that PKF vaccination plan induced a polarized Th1 response and rendered an effective protection against bloodstream infection caused by A. baumannii. • In vivo immunogenic reactivity of PKF, AbOmpA and PKF + AbOmpA were evaluated against sepsis of Acinetobacter baumannii. • High concentration of Total-IgG and IgG2c and low concentration of IgG1 were detected in all immunization plans. • PKF immunization plan fulfilled a more efficient protection against sepsis of A. baumannii. [ABSTRACT FROM AUTHOR]

Marek's disease virus (MDV) is a cell-associated α-herpesvirus of chickens. It is difficult to grow MDV in suspension culture. Therefore, MDV vaccines are currently produced using adherent primary chicken embryo fibroblasts, and on a large scale this is labour-intensive and costly. In this study, the CVI988 strain of MDV was inoculated into chicken fibroblast cell line UMNSAH/DF-1 (DF-1) cultured by microcarrier suspension for the proliferation experiment. Moreover, the effects of culture conditions, such as inoculation method, multiplicity of infection (MOI), microcarrier concentration, and pH value, on the proliferation of MDV were investigated. The results demonstrated that the maximum viral load of 64.76 ± 2.64 × 106 PFU/flask in a working volume of 100 ml could be obtained using synchronous cell seeding and inoculation method at an MOI of 0.02 and a microcarrier concentration of 5 g/l at pH 7.2. At the same time, the CVI988/DF-1 vaccines prepared by the microcarrier culture process and the traditional adherent cell culture process (CVI988/Rispens) were compared through bird experiments. We found a protective rate of 94.4% using the CVI988/DF-1 vaccine with specific pathogen-free chickens that was equivalent to that of the commercial vaccine CVI988/Rispens (protection rate of 94.1%). In this study, the MDV CVI988/DF-1 vaccine prepared by the microcarrier suspension culture of DF-1 cells could provide effective immune protection for specific pathogen-free chickens, providing a reference for the prevention and control of MD and further development of a large-scale bioreactor for producing the MD vaccine. [ABSTRACT FROM AUTHOR]

High temperature is known to cause some instability in polysaccharide-protein conjugated vaccines and studies under stress conditions may be useful in determining whether short-term accidental exposure to undesired conditions can compromise product quality. In this study, we examined the structural stability of three industrial batches of Brazilian Meningococcal C conjugate bulk (MPCT) incubated at 4, 37, and 55 °C for 5 weeks. The effect of exposure to the storage temperatures was monitored by HPLC-SEC, CZE, CD and NMR techniques. The immunological significance of any physicochemical changes observed in MPCT was determined by SBA and ELISA assays of serum from immunized mice. Fluorescence emission spectra at 4 and 37 °C were similar among all samples and compatible with the native fold of the carrier protein. Fluorescence spectra of MPCT stored at 55 °C decreased in intensity and had a significant red-shift, indicating conformational changes. Far-UV CD spectra revealed a trend toward loss of structural conformation as storage temperature was increased to 55 °C. The NMR data showed modified signal intensity of the aromatic and aliphatic residues, mainly for samples incubated at 55 °C, suggesting a partial loss of tertiary structure. About 50% free saccharide content was found in bulks stored at 55 °C, but no difference was observed in the IgG or SBA titers. The present study showed physicochemical methods alone are insufficient to predict the biological activity of a MPCT conjugate vaccine without extensive validation against immunological data. However, they provide a sensitive means of detecting changes induced in a vaccine exposed to adverse environmental condition. [ABSTRACT FROM AUTHOR]

Abstract. Members of the European Society for Immunodeficiencies (ESID) and other colleagues have updated themulti-stage expert-opinion-based diagnostic protocol for non-immunologists incorporating newly defined primary immunodeficiency diseases (PIDs). The protocol presented here aims to increase the awareness of PIDs among doctors working in different fields. Prompt identification of PID is important for prognosis, but this may not be an easy task. The protocol therefore starts from the clinical presentation of the patient. Because PIDs may present at all ages, this protocol is aimed at both adult and paediatric physicians. The multi-stage design allows cost-effective screening for PID of the large number of potential cases in the early phases, with more expensive tests reserved for definitive classification in collaboration with a specialist in the field of immunodeficiency at a later stage.

Especially complicated, renal lithiasis contributes to the general inflammatory syndrome development that interferes with nonspecific, humoral and cellular immune system. The surgical treatment of nephrolithiasis is closely related to drug therapy of urinary infection, one of the reasons being the reduction of the immune status. The work is performed by evaluating the immunological status preoperatively in 58 patients with complicated lithiasis. The analysis of the status in these patients demonstrated that complicated urolithiasis results in significant changes in the immune system, these changes being expressed at the cellular and humoral level of immunity. [ABSTRACT FROM AUTHOR]

Crimean-Congo hemorrhagic fever (CCHF) is a zoonotic disease caused by Crimean-Congo hemorrhagic fever virus (CCHFV), which can cause severe clinical disease and even death in humans. In recent years, the disease has spread to a wider area, posing a major public health threat to China as well as the Middle East, Europe and Africa, and there is no safe and effective vaccine to prevent the disease. Recently, it has been shown that using the Zera fusion to target proteins can enhance immunogenicity and improve the potential for developing viral vaccines. Based on this finding, in this study, two vaccine candidates, Zera-Gn and Zera-Np, were prepared using an insect baculovirus system expressing CCHFV glycoprotein (Gn) and nucleocapsid protein (Np) fused with Zera tags, and evaluated for immunogenicity in BALB/c mice. The obtainedresults showed that both Zera-Gn and Zera-Np recombinant nanoparticles were successfully expressed, and Zera-Gn had good induction of humoral and cellular immunity in mice, and its immunogenicity was significantly higher than that of Zera-Np. The results indicated that Zera-Gn self-assembled nanoparticles prepared by fusing Zera tags with CCHFV spike-in protein Gn have the potential to be a candidate vaccine for CCHF, and this study provides a reference for the development of Zera self-assembled nanoparticle vaccine for CCHF., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Zhang, Wang, Jiang, Kong, Wang, Li and Zhang.)

Antibody deficiency is common finding in patients with Jacobsen syndrome (JS). In addition, there have been few reports of T-cell defects in this condition, possibly because most of the reported patients have not been specifically evaluated for T-cell function. In this article, we present a child with an 11q deletion and combined immunodeficiency and we perform a literature overview on immunodeficiency in JS. Our patient presented with recurrent bacterial and prolonged viral infections involving the respiratory system, as well as other classic features of the syndrome. In addition to low IgM, IgG4, and B-cells, also low recent thymic emigrants, helper and naïve T-cells were found. We propose that patients with Jacobsen syndrome need thorough immunological evaluations as T-cell dysfunction might be more prevalent than previously reported. Patients with infections consistent with T-cell defects should be classified as having combined immunodeficiency. [ABSTRACT FROM AUTHOR]

Crimean-Congo hemorrhagic fever (CCHF), which has a fatality rate of 20-30%, is widely prevalent in several regions in Asia, Europe, and Africa and has spread to a wider range of areas in recent years. At present, there is a lack of safe and effective vaccines for the prevention of CCHF. In this study, we prepared three vaccine candidates, rvAc-Gn, rvAc-Np, and rvAc-Gn-Np, that encoded the CCHF virus (CCHFV) glycoprotein Gn and the nucleocapsid protein (Np) on the surface of baculovirus using an insect baculovirus vector expression system (BVES) and evaluated their immunogenicity in BALB/c mice. The experimental results showed that both CCHFV Gn and Np were expressed by the respective recombinant baculoviruses and anchored to the viral envelope. BALB/c mice were immunized, and all three recombinant baculoviruses showed significant humoral immunity. At the cellular level, the level of immunity in the rvAc-Gn group was significantly higher than that in the rvAc-Np and rvAc-Gn-Np groups, and the rvAc-Gn-Np coexpression group exhibited the lowest level of cellular immunity. In conclusion, the strategy of coexpressing Gn and Np in the baculovirus surface display system did not result in improvements in immunogenicity, whereas the recombinant baculovirus displaying Gn alone could induce significant humoral and cellular immunity in mice, indicating that rvAc-Gn has potential as a CCHF vaccine candidate. This study thus provides new ideas for the development of a CCHF baculovirus vaccine., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Zhang, Wang, Jiang, Wang, Zhang and Li.)

Objectives To present some immunological aspects of fresh-frozen allogeneic bone grafting for lateral bone augmentation, based on the quantitative evaluation of IL-10, IL-1β, IFN- γ and TNF- α in patients sera. Material and methods Thirty-three partially or totally edentulous patients received fresh-frozen allogeneic bone ( AL - 20 patients) or autologous bone onlay block grafts ( AT - 13 patients) prior to oral implant placement. Blood samples were collected from each patient at various time-points during a 6 month-period (baseline, 14, 30, 90 and 180 days postoperatively). Quantitative evaluation of IL-10, IL-1β, IFN- γ and TNF- α was performed by enzyme linked immunosorbent assay ( ELISA). Results For all evaluated markers and at all evaluated periods, inter-group comparisons showed no statistically significant differences between the groups, while the observed values were within normal levels. For AL-treated patients, intra-group evaluation showed statistically significant increase of TNF-α from baseline to 90 ( P < 0.001) and 180 ( P < 0.01) days, and from 14 to 90 ( P < 0.01) and 180 ( P < 0.05) days. IFN- γ showed intercalated results, with a decrease from baseline to 14 days ( P < 0.05), and increase from 14 to 90 days ( P < 0.001) and 180 ( P < 0.05) days. No differences between the periods of evaluation were found for the AT group. Conclusions AL grafting for lateral bone augmentation, similar to AT grafting, does not seem to challenge the immune system significantly. [ABSTRACT FROM AUTHOR]

Abstract: Combined vaccines are immunological products intended for immunization against multifactorial infectious diseases caused by different types or variants of pathogens. In this study, the effectiveness of Vibrio alginolyticus (Va), Vibrio harveyi (Vh), Vibrio vulnificus (Vv) and infectious spleen and kidney necrosis virus (ISKNV), an iridovirus, combined-vaccine (Vibrio and ISKNV combined vaccines, VICV), Va+Vh+Vv inactive vaccine (VIV) and ISKNV whole cell inactive vaccine (IWCIV) in Epinephelus coioides were evaluated using various immunological parameters including antibody titer, serum lysozyme activity (LA), respiratory burst (RB) activity, bactericidal activity (BA) and relative percentage survival (RPS). E. coioides immunized with VICV and challenged with Va+Vh+Vv+ISKNV had an RPS of 80%. The RPS was 73.3% in E. coioides immunized with VIV and challenged with Va+Vh+Vv. E. coioides immunized with IWCIV and challenged with ISKNV had an RPS of 69.6%. Serum LA in the vaccinated group was significantly higher than the control group on days 21 and 28 post-vaccination (P <0.01). The RB activity of head kidney cells in the vaccinated group was significantly higher (P <0.01) compared to that in the control group. However, RB activity of spleen cells in the vaccinated group and the control group were not significantly different (P >0.05). After immunization with VICV, BA values of blood leucocytes and head kidney cells increased significantly more than spleen cells. BA value of blood leucocytes was higher than that in head kidney cells. There were distinct difference between BA values in head kidney cells and in spleen cells (P <0.05) as well as between BA value of blood leucocytes and head kidney cells (P <0.01). E. coioides vaccinated with VICV have significantly higher antibody levels than control groupers (P <0.01). Our study suggests that the VICV candidate can effectively protect groupers against multiple bacterial and viral pathogens. [Copyright &y& Elsevier]

Natural polymer like potato starch is a mixture of amylose and amylopectin, . Extraction of Potato starch was performed by rasping, centrifugation, refining and drying method. In our research method, we employed potato starch as a biodegradable polymer; It has a great impact on pharmaceutical applications due to its bioavailability, non toxic, high change density and biodegradability. In our research work, we have selected Potato starch polymer as a model of immunomodulatory effect of vaccine of tetanus antigen. According to WHO, Tetanus is a systemic infectious disease caused by genus Cl Tetanii. It has been estimated that the tetanus fever endemicity among large populations and global emergence of multidrug resistant strains to impose greater urgency on the evaluation of existing and new vaccines to prevent mortality of neonates and in pregnancy. Recently available recombinant vaccine was seems to be side effect and cost effective. The starch polymer in the form of microspheres was preferred in order to replace the alum to elicit sustained immune response because alum induces local granuloma and hypersensitivity reaction to some individual. We have employed microencapsulation technique by using 0.5% ml glutaraldehyde as a crosslinking agent. The particle size was analyzed as 40.23μm. Invitro studies was analyzed by SEM, stabilities studies Immunogenicity studies was carried out by incubating the sample, centrifuged and tested for an antigen and the Compatibility study was performed by Infrared Spectroscopy, the antigen integrity was studied by SDS PAGE and ELISA. Immunoglobulin titer values was found out ( IgG, IgA,IgM, IgE) to show the increase level of antibody response. [ABSTRACT FROM AUTHOR]

Efficient early identification of primary immunodeficiency disease (PID) is important for prognosis, but is not an easy task for non-immunologists. The Clinical Working Party of the European Society for Immunodeficiencies (ESID) has composed a multi-stage diagnostic protocol that is based on expert opinion, in order to increase the awareness of PID among doctors working in different fields. The protocol starts from the clinical presentation of the patient; immunological skills are not needed for its use. The multi-stage design allows cost-effective screening for PID within the large pool of potential cases in all hospitals in the early phases, while more expensive tests are reserved for definitive classification in collaboration with an immunologist at a later stage. Although many PIDs present in childhood, others may present at any age. The protocols presented here are therefore aimed at both adult physicians and paediatricians. While designed for use throughout Europe, there will be national differences which may make modification of this generic algorithm necessary. [ABSTRACT FROM AUTHOR]

Background: Tuberculosis (TB) is an infectious disease that poses a significant health threat and is one of the leading causes of death worldwide. Diabetes mellitus (DM) has high morbidity and mortality rates. Previous studies have reported that comorbidities can influence one another and aggravate immune disorders. A systematic and comprehensive evaluation of the immune status of patients with TB and DM (TB-DM) is helpful for early clinical immune intervention and for promoting the recovery of patients with TB-DM., Methods: This study included 159 patients with TB without DM (TB-NDM) and 168 patients with TB-DM. Interferon-γ (IFN-γ) release assays (IGRAs) and TB-specific antibodies against 38kD+16kD proteins were used to detect humoral and cellular immune responses. Flow cytometry was used to analyze the absolute counts of the lymphocyte subsets., Results: There was no significant difference in the positive rate of enzyme-linked immunospot (ELISPOT) assays, enzyme linked immunosorbent assay (ELISA), and 38kD+16kD antibodies between the TB-DM and TB-NDM groups. Pulmonary lobe lesion and cavity formation rates were significantly higher in patients with TB-DM with poor glycemic control than patients with TB-NDM and TB-DM with normal glycemic control. The absolute counts of T lymphocytes, CD8+ T lymphocytes, and B lymphocytes in patients with TB-DM were markedly lower than those in patients with TB-NDM. The absolute counts of T lymphocytes and CD8+ T lymphocytes in patients with TB-DM and hyperglycemia were lower than those in patients with euglycemia. Linear regression analysis revealed that the absolute counts of total T lymphocytes, CD8+ T lymphocytes, and NK cells in patients with TB-DM significantly decreased with increasing fasting blood glucose (FBG) levels., Conclusion: Hyperglycemia is a risk factor for pulmonary cavity formation and lobe lesions in patients with TB-DM and suppresses the absolute counts of total T lymphocytes, CD8+ T lymphocytes, and NK cells in patients with TB-DM. The potential mechanism may involve the downregulation of innate and adaptive immune responses., Competing Interests: The authors declare no conflicts of interest in this work., (© 2022 Wei et al.)

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Pertussis is an acute respiratory tract infection caused by Bordetella pertussis. Even though its current vaccine coverage is relatively broad, they still have some shortcomings such as short protection time and might be incapable of blocking the spread of the disease. In this study, we developed new pertussis vaccine candidates by separately fusing three pertussis antigens (B. pertussis fimbriae 2 "Fim2", pertussis toxin S1 subunit "PtxS1", and filamentous hemagglutinin "FHA1877–2250") to each of two immune-boosting carrier proteins (B subunits of AB5 toxin family: cholera toxin B subunit "CTB" and shiga toxin B subunit "StxB"). We then immunized mice with these fusion antigens and found that they significantly increased the serum antibody titers and elicited high bactericidal activity against B. pertussis. After CTB-or StxB-fused antigen-immunized mice were challenged with a non-lethal dose of B. pertussis, the bacterial loads in different tissues of these mice were significantly reduced, and their lung damage was nearly invisible. Furthermore, we also demonstrated that these candidate vaccines could provide strong prophylactic effects against a lethal challenge with B. pertussis. Overall, our candidate vaccines conferred better immune protection to mice compared with pertussis antigen alone. This B5 subunit-based vaccine strategy provides a promising option for vaccine design. [ABSTRACT FROM AUTHOR]

Objectives: Inborn Errors of Immunity are characterized by infectious conditions and manifestations of immune dysregulation. The diversity of clinical phenotypes can make it difficult to direct the laboratory investigation. This article aims to update the investigation of immunological competence in the context of primary defects of the immune system., Source of Data: Searches were carried out on Pubmed to review articles published in the last five years, in English, French or Spanish, using the terms "diagnosis" OR "investigation" AND "immunodeficiency" or "primary immunodeficiency" or "inborn errors of immunity" NOT "HIV". Recent textbook editions have also been consulted., Summary of Findings: The immune system competence investigation should be started based on clinical phenotypes. Relevant data are: characterization of infectious conditions (location, recurrence, types of infectious agents, response to treatment), age during symptom onset and associated manifestations (growth impairment, allergy, autoimmunity, malignancies, fever and signs of inflammation without the identification of infection or autoimmunity) and family history. These data contribute to the selection of tests to be performed., Conclusions: The diagnostic investigation of Inborn Errors of Immunity should be guided by the clinical characterization of patients, aiming to optimize the use of complementary tests. Many diagnoses are attained only through genetic tests, which are not always available. However, the absence of a diagnosis of certainty should never delay the implementation of therapeutic measures that preserve patient life and health., (Copyright © 2020 Sociedade Brasileira de Pediatria. Published by Elsevier Editora Ltda. All rights reserved.)

Background: Several approaches have not been successful to suppress HIV (Human immunodeficiency virus) infection among infected individuals or to prevent it yet. In order to expand strong HIV specific humoral and cellular responses, Virus-like particles (VLPs) as potential vaccines show significant increase in neutralizing antibodies secretion, T-cell count and also secretion of cytokines., Objective: This study aimed at immunological evaluation of VLPs harboring high copy of MPERV3 in BALB/c mice., Methods: Female BALB/c mice were immunized with homologous and heterologous primeboosting regimens of HIV-1 VLPMPER-V3. Their immune responses were evaluated for humoral responses (Total IgG and IgG isotyping) and cellular responses (IFN-γ, IL-5 secretion, in vitro CTL assay and T cell proliferation) and compared in immunized mice., Results: The data showed robust induction of humoral response in mice groups which received different regimens of VLP. Furthermore, analysis of cytokine profile indicated that the highest IL-5 secretion was related to VLP+M50 group and confirmed the dominance of Th2 immunity in this group., Conclusion: This study showed that VLP MPER-V3 as a potential vaccine candidate has the potency as an effective prophylactic vaccine and this finding guarantees further investigations to achieve a promising HIV-1 vaccine candidate., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)