Increased IgG Levels Research Articles

First real-world experience of Gam-COVID-Vac Sputnik V vaccine use

With the ongoing SARS-CoV-2 pandemic, the safety and efficacy of vaccine prophylaxis with new vaccines have become vital. Objective. To study tolerability and efficacy of the Gam-COVID-Vac (Sputnik V) vaccine in the short term in a real-world setting in the example of the Federal remote consulting center on vaccination against new coronavirus infection COVID-19 of National Medical Research Center for Therapy and Preventive Medicine. Materials and methods. We retrospectively analyzed data from the registration cards of the first 500 voluntary vaccination recipients who received two vaccine doses in October-December 2020. Serum immunoglobulin (Ig) M and G levels to SARS-CoV-2 were measured in 204 participants at baseline, at Day 21, and at Day 42 after the injection of component I of Gam-COVID-Vac vaccine. Test for SARS-CoV-2 coronavirus RNA in a nasopharyngeal and/or oropharyngeal mucosa smear was performed before injection of component I of the vaccine using real-time PCR method on a DT-96 amplifier (DNA-Technology, Russia) using SARSCoV-2/SARS-CoV reagent kit (DNA-Technology, Russia). Anti-SARS-CoV-2 IgG and IgM were measured using reagent kits for enzyme immunoassay (Vector-Best, Russia). Results. During the follow-up (42 days), none of the vaccine recipients experienced serious adverse events. During the first 14 days after receiving component I of the vaccine Gam-COVID-Vac, eight vaccine recipients got sick with COVID-19;the disease had a mild course and did not require hospital admission. On Day 21, no IgM antibodies were detected in any of the vaccine recipients, while the IgG level was detectable and the cutoff index (CI) was 11.3 [3.4;16.2]. On Day 42 after the injection of component I of the Gam-COVID-Vac vaccine, there was an increase in IgG levels, and CI was 16.2 [16.0;16.4] in most subjects, while no IgM levels were detected in any of the vaccine recipients tested. Conclusion. The data obtained may indicate a favorable tolerability and safety profile of the combined vector vaccine Gam-COVID-Vac, as well as short-term efficacy, according to the change of anti-SARS-CoV-2 IgG and absence of severe COVID-19 disease in vaccinated subjects during the follow-up period. To obtain further evidence of the long-term clinical efficacy of Gam-COVID-Vac, follow-up of vaccinated subjects should be continued. (English) [ABSTRACT FROM AUTHOR] Ð’ NƒN

TDP-43-specific Autoantibody Decline in Patients With Amyotrophic Lateral Sclerosis.

ObjectiveWe hypothesize alterations in the quality and quantity of anti–43-kDa TAR DNA-binding protein (TDP-43) naturally occurring autoantibodies (NAbs) in patients with amyotrophic lateral sclerosis (ALS); therefore, we assessed relative binding properties of anti–TDP-43 NAbs composite in plasma from patients with ALS in comparison with healthy individuals.MethodsELISA competition assay was used to explore the apparent avidity/affinity of anti–TDP-43 NAbs in plasma from 51 normal controls and 30 patients with ALS. Furthermore, the relative levels of anti–TDP-43 NAbs within the immunoglobulin (Ig) classes of IgG (isotype IgG1-4) and IgMs were measured using classical indirect ELISA. The occurring results were hereafter correlated with the measures of disease duration and disease progression.ResultsHigh-avidity/affinity anti–TDP-43 NAbs levels were significantly reduced in plasma samples from patients with ALS. In addition, a significant decrease in relative levels of anti–TDP-43 IgG3 and IgM NAbs and a significant increase in anti–TDP-43 IgG4 NAbs were observed in ALS plasma vs controls. Furthermore, a decrease in global IgM and an increase in IgG4 levels were observed in ALS. These aberrations of humoral immunity correlated with disease duration, but did not correlate with ALS Functional Rating Scale–Revised scores.ConclusionsOur results may suggest TDP-43–specific immune aberrations in patients with ALS. The skewed immune profiles observed in patients with ALS could indicate a deficiency in the clearance capacity and/or blocking of TDP-43 transmission and propagation. The decrease in levels of high affinity/avidity anti-TDP-43 NAbs and IgMs correlates with disease progression and may be disease predictors.

The immune gene expression induced by Cooperia punctata in naturally infected calves with resistance and susceptible phenotype traits

The analysis of the immune gene expression was performed in Zebu × Holstein calves with resistant and susceptible phenotypes naturally infected with Cooperia punctata. Fourteen calves of 4 months old were grazed for 11 weeks under a tropical climate. The parasitic infection showed an average epg value of 1055 ± 1155 and an IgG optical density of 0.814 ± 0.0.037 with statistic differences among the different weeks (p < 0.05), and a pcv value of 24 ± 2.0 % (p > 0.05). High variation in epg value was observed, between 7 ± 7.14 and 4657 ± 1886, and, based on these differences; the infected hosts were classified as five resistant calves with epg ≤ 200 and nine susceptible calves with epg ≥ 300. Moreover, IgG levels displayed statistical differences between resistance and susceptible calves to C. punctata infection. The immune gene expression was analysed in three resistant and susceptible calves, respectively. Nine cytokine genes and the FCεR1A receptor were analysed at the 3rd and 11th weeks post-infection. In the first period upregulation was found, from 2.19- to 9.45-fold, (p < 0.05) for IL-2, -5, - 6, -10, TGF-β and FCεR1A in the resistant group; the expression was decreased at the 11th week with low level of IgG. In contrast, downregulation for susceptible calves was found for nine immune genes and upregulation for INF-γ in both periods together with increased IgG levels. In conclusion, immune gene expression was regulated at the begging infection of C. punctata in resistant grazing calves. In contrast, suppression of important genes was involved in calves susceptible to C. punctata.

Gestational Dysfunction-Driven Diets and Probiotic Supplementation Correlate with the Profile of Allergen-Specific Antibodies in the Serum of Allergy Sufferers.

Background: Maternal diet has significant effects on development of childhood atopic disease and hypersensitivity development. However, the gestational dysfunctions demanding special diets are becoming a widespread phenomenon, their immunological implications can be manifested in the profile of antibodies in the offspring’s serum. Methods: 153 allergic and 150 healthy individuals were diagnosed for allergy using specific antibody and cytokine immunoassay tests. The medical history of subjects along with mothers’ course of pregnancy was completed by allergologist’s anamnesis. A self-organizing neural network and multivariate analyses to complex data and pick basic interactions were used. Results: Two significant explanatory modules were determined. The first was formed by gestational diabetic and cholestatic diet, infant formula feeding type, probiotic supplementation and its BMI index, moderate IgE, increased IgG levels of antibodies and single or poly-food allergy type (7 clusters). The second was formed by gestational vegan/vegetarian and elimination diet, maternal probiotic supplementation, sex, high IgE total antibodies and food and mixed poly-allergy to aero- and food-origin allergens (19 clusters). Conclusions: Significant associations were observed between special gestational diet intake underlying foetal programming and the mechanisms of childhood allergy. The novelty is the positive association between diabetic and cholestatic diet intake and IgE/IgG-mediated food hypersensitivity.

Extraction and biological evaluation of external membrane vesicles of Brucella abortus as a candidate for brucellosis vaccine

Amaç: Bruselloz, hayvan ve insan sağlığını etkileyen endemik bir zoonotik hastalıktır. Son birkaç on yılda, hastalığı kontrol etmek için daha güvenli Brucella aşıları geliştirmek için çok araştırma yapılmıştır. Dış zar vezikülleri (OMV'ler), insan brusellozunu önlemek için bir alt birim aşı için immünojenik yapılar olarak düşünülmüştür. Bu çalışmanın amacı Brusella abortus'un dış zar veziküllerini bruselloz aşısı için aday olarak değerlendirmektir.Yöntemler: Bu çalışmada Brucella abortus, S99 suşu kullanıldı. Brucella abortusun kitle kültivasyonundan sonra OMV ekstraksiyonu, Ultracentrifugation ve Sodium Dosoxycotinate tarafından gerçekleştirildi. SDS-PAGE yöntemi, fizikokimyasal özellikleri değerlendirmek için protein paternini ve elektron mikroskopisini gözlemlemek için kullanıldı. OMV'deki LOS miktarı da LAL testi ile ölçüldü. Protein konsantrasyonunun belirlenmesinden sonra farelere belirli bir konsantrasyonda enjekte edildi ve antikoru değerlendirmek için kan örnekleri alındı. Son olarak serum izolasyonundan sonra antikor üretimi ELISA yöntemi ile ölçüldü.Bulgular: OMV'de mevcut protein miktarı 0,1 mg/ml idi. OMV'nin boyutu 55 ila 150 nm idi ve SDS-PAGE değerlendirmesinde protein aralığı >25 kDa idi. LAL testindeki LOS değeri yetkili aralıkta rapor edildi. İlk enjeksiyondan sonra IgG düzeylerinde (P=0,001) kontrol grubuna kıyasla ve daha sonraki güçlendiricilerde anlamlı artış gözlendi. En yüksek tepki oranı ikinci güçlendiricide gözlenmiştir.Sonuç: OMV ekstraksiyon sırasında uzamsal şeklini korumaktadır ve Brucella düşüklerine karşı yüksek derecede spesifik antikorların üretimini indükleme potansiyeline sahiptir. Bu nedenle klinik fazları inceledikten sonra aday bir aşı olarak düşünülebilir.

Immune status in sarcoidosis: one size does not fit all.

B-cell immunity and immunoglobulins are less commonly affected in sarcoidosis. We aimed to evaluate immune status in sarcoidosis patients. Retrospective chart review of sarcoidosis patients attending a out-patient clinic over 3 months period. Immunoglobulins levels were recorded (A, M, G, E) along with clinical and serological data. They were divided in group A (normal IgG), group B (increased IgG), group C (decreased IgG) and group D (decreased IgG and IgM and/or IgA). Of 50 subjects, 68% were females and 62% of Caucasian origin. 22 (44%) had normal IgG levels, 16 (32%) had increased IgG levels, 10 (20%) had hypogammaglobulinemia and 2 (4%) had combined hypogammaglobulinemia, diagnosed with combined sarcoidosis and common variable immunodeficiency. Decreased IgA values was found in groups C and D. IgE was high in group B. Globulin was increased in group B and decreased in groups C and D. Decreased neutrophils were found in group D (all statistically significant). Correlation analysis showed significant association of angiotensin converting enzyme with IgA and IgM, inverse correlation of IgG with white blood cells and neutrophils, of IgA with globulin and inverse with albumin and of calcium with albumin. Most sarcoidosis patients have normal or increased immunoglobulin levels, that correlate with serum biomarkers of disease activity. Hypogammaglobulinemia may reflect treatment side effects and accompanied by blood leukocytosis. Combined severe immunodeficiency occurs in sarcoidosis.

Antibodies to Epstein-Barr virus and neurotropic viruses in multiple sclerosis and optic neuritis

Antibody indices to Measles, Mumps, Varicella Zoster (MRZ) are of diagnostic value in multiple sclerosis (MS). Here, we have investigated, if this panel could be extended to increase diagnostic value.Samples from relapsing-remitting (RR) MS and optic neuritis (ON) patients were tested for reactivity to antigens from Epstein-Barr, Varicella Zoster, Measles, Mumps and Rubella (EMMRZ) viruses. Increased IgG levels in serum and cerebrospinal fluid (CSF) were found in RRMS patients, along with a significant correlation between serum and CSF. The sensitivity of the EMMRZ panel was increased approximately 40% compared to the MRZ panel, suggesting that the EMMRZ panel may be useful in MS and ON diagnostics.

Various surgical techniques influence on the local immunity state in the postoperative period in children's tonsils hypertrophy

Topicality: There are many conflicting opinions in the literature regarding the best surgical technique for the treatment of OSAS related to palatine hypertrophy – partial (tonsillotomy) or complete (tonsillectomy) removal of lymphoid tissue of the tonsils. Also, there are many opinions about the use of a variety of traditional and modern surgical instruments to reduce the volume of the tonsils, including using a laser, a radiofrequency knife, an ultrasonic knife, a shaver (microdebrider) and a coblator. Aim of the study: The aim of the study was to compare the surgical methods effectiveness of tonsils hypertrophy treatment which the based on the restoration of the mucous membrane's local immunity function. Materials and methods: Children who undergoing surgical treatment for palatine tonsils hypertrophy 2-3 degrees according to the Brodsky classification and the presence of OSAS took part in the study. The patients were randomized and divided into four comparison groups using different surgical techniques. The first group, which used the surgical technique of classical tonsillotomy (TT) included 28 patients, the second group, which used the surgical technique of intracapsular shavertonsillotomy(STT) included 18 patients, the third group, which used surgical technique of intracapsular coblationtonsillotomy (CTT), and groups which using surgical technique of tonsilloplasty included 12 patients (TP). Determined the state of local immunity before surgery and 1 month after surgery by concentration levels of sIgA, α-IFN, IL-1β, TGF-1β and general-purpose immune complexes. Results: Recovery of the concentration of the major marker of local immunity sIgA occurs significantly (P&lt;0.05) faster when using the surgical technique of CTT and STT. Recovery of the concentration of the major marker of antiviral immunity α-IFN occurs significantly (P &lt;0.05) faster when using the surgical technique of CTT, STT and TP. Recovery of the concentration of the anti-inflammatory cytokine IL-1β occurs significantly (P&lt;0.05) faster with the use of the surgical technique of STT and TP. Indicators of the regeneration factor TGF-1β are significantly the highest when using the surgical techniques of STT, TT and TP. The concentration of immune complexes in the oropharyngeal secretions of patients with tonsil hypertrophy at 1 month after surgery significantly decreased with the use of surgical techniques TT and TP. Significant (P &lt;0.05) increase in IgG level was de- termined in all comparison groups except the group where tonsilloplasty was used. Conclusions: The safety surgical technique of palatine tonsils volume reduction – TP and STT promotes significantly (P&lt;0,05) faster restoration of concentration of the main mucous membrane's local immunity markers.

Immunomodulatory effect of papaya (Carica papaya) pulp and seed extracts as a potential natural treatment for bacterial stress.

The current study evaluated the immunomodulatory effects of Carica papaya pulp and seeds methanol (MeOH) extracts on mice infected with Listeria monocytogenes. Gas chromatography-mass spectrometry analysis identified 10 active constituents in C. papaya seed MeOH extract and 10 compounds in C. papaya pulp MeOH extract. The experimental animals were divided into negative control (G1) group, positive control (G2) group, pulp extract treated (G3) group, and seed extract treated (G4) group. After infection of animals (G2, G3, and G4), treatments were started for 3weeks. Estimation of the immunological parameters showed a marked decrease in IgM levels and an increase in IgG levels in the treated groups (G3 and G4) compared with those in G2. The proinflammatory cytokines (IL-10, IL-12, IL-1β, IL-6, and TGF-β1) were decreased in the treated groups (G3 and G4) compared with those in G2. Nitric oxide levels were also decreased, and the percentages of phagocytosis increased compared with those of G2. The results demonstrated the immunomodulatory and anti-inflammatory effects of C. papaya pulp and seeds MeOH extracts. PRACTICAL APPLICATIONS: Based on the antioxidant and antibacterial activities exhibited by the pulp and seed MeOH extracts investigated in this study, Carica papaya might be considered as a natural source of phytochemicals that could be utilized in novel foods and pharmaceuticals. Further investigation are needed to identify and purify compounds that might be responsible for the observed effects.

JC polyomavirus-specific antibody responses in pediatric kidney transplant recipients.

BKPyV is widely recognized in KTRs, but little is known about rates of primary and secondary JCPyV exposure in pediatric KTRs. Weevaluated JCPyV exposure in pediatric KTRs using antibody responses in the first 12months post-transplant. Of 46 children transplanted between 2009 and 2014, 6 lacked any samples for serologic testing, leaving 40 KTRs for study. JCPyV-specific IgG and IgM antibodies were measured using a normalized VLP ELISA. Significant JCPyV exposure was defined as IgG seroconversion, increasing IgG levels of >0.5 nOD units, or IgM detection. Of 40 recipients (median age 3.2years), 11 (27.5%) were seropositive, 20 (50%) seronegative for JCPyV-IgG, while 9 (22.5%) had no specimen at the time of transplantation, but were confirmed as seronegative in post-transplant samples. Of 29 (72.5%) at risk, JCPyV-IgG seroconversion occurred in 15/29 (51.7%) including JCPyV-IgM in 6 patients (20.7%). Two patients (6.9%) developed only JCPyV-IgM. Among JCPyV-IgG-positive KTRs, six (12.5%) had significant IgG increases. Altogether 23 of 40 patients (57.5%) had serological evidence of primary or secondary JCPyV exposure. In these patients, kidney function tended to be lower during the 2years of follow-up, but only one patient lost the graft due to JCPyV nephropathy.Thus, JCPyV exposure is common in pediatric KTR and may present serologically as primary or secondary infection. Although only one case of JC-PyVAN occurred, a trend toward lower renal function was seen. Dedicated studies of larger cohorts are warranted to define impact of JCPyV in pediatric KTR.

RTS,S/AS01E immunization increases antibody responses to vaccine-unrelated Plasmodium falciparum antigens associated with protection against clinical malaria in African children: a case-control study

BackgroundVaccination and naturally acquired immunity against microbial pathogens may have complex interactions that influence disease outcomes. To date, only vaccine-specific immune responses have routinely been investigated in malaria vaccine trials conducted in endemic areas. We hypothesized that RTS,S/A01E immunization affects acquisition of antibodies to Plasmodium falciparum antigens not included in the vaccine and that such responses have an impact on overall malaria protective immunity.MethodsWe evaluated IgM and IgG responses to 38 P. falciparum proteins putatively involved in naturally acquired immunity to malaria in 195 young children participating in a case-control study nested within the African phase 3 clinical trial of RTS,S/AS01E (MAL055 NCT00866619) in two sites of different transmission intensity (Kintampo high and Manhiça moderate/low). We measured antibody levels by quantitative suspension array technology and applied regression models, multimarker analysis, and machine learning techniques to analyze factors affecting their levels and correlates of protection.ResultsRTS,S/AS01E immunization decreased antibody responses to parasite antigens considered as markers of exposure (MSP142, AMA1) and levels correlated with risk of clinical malaria over 1-year follow-up. In addition, we show for the first time that RTS,S vaccination increased IgG levels to a specific group of pre-erythrocytic and blood-stage antigens (MSP5, MSP1 block 2, RH4.2, EBA140, and SSP2/TRAP) which levels correlated with protection against clinical malaria (odds ratio [95% confidence interval] 0.53 [0.3–0.93], p = 0.03, for MSP1; 0.52 [0.26–0.98], p = 0.05, for SSP2) in multivariable logistic regression analyses.ConclusionsIncreased antibody responses to specific P. falciparum antigens in subjects immunized with this partially efficacious vaccine upon natural infection may contribute to overall protective immunity against malaria. Inclusion of such antigens in multivalent constructs could result in more efficacious second-generation multistage vaccines.

Concentration and avidity of antibodies to different circumsporozoite epitopes correlate with RTS,S/AS01E malaria vaccine efficacy

RTS,S/AS01E has been tested in a phase 3 malaria vaccine study with partial efficacy in African children and infants. In a cohort of 1028 subjects from one low (Bagomoyo) and two high (Nanoro, Kintampo) malaria transmission sites, we analysed IgG plasma/serum concentration and avidity to CSP (NANP-repeat and C-terminal domains) after a 3-dose vaccination against time to clinical malaria events during 12-months. Here we report that RTS,S/AS01E induces substantial increases in IgG levels from pre- to post-vaccination (p < 0.001), higher in NANP than C-terminus (2855 vs 1297 proportional change between means), and higher concentrations and avidities in children than infants (p < 0.001). Baseline CSP IgG levels are elevated in malaria cases than controls (p < 0.001). Both, IgG magnitude to NANP (hazard ratio [95% confidence interval] 0.61 [0.48–0.76]) and avidity to C-terminus (0.07 [0.05–0.90]) post-vaccination are significantly associated with vaccine efficacy. IgG avidity to the C-terminus emerges as a significant contributor to RTS,S/AS01E-mediated protection.

Immunomodulatory effects of Alhagi honey polysaccharides encapsulated into PLGA nanoparticles

Alhagi honey polysaccharides (AHP) have been widely studied as immunomodulators. Poly(lactic-co-glycolic acid) (PLGA) nanoparticles have been frequently used to control the release of drugs. In this study, AHP was extracted and encapsulated within PLGA (AHPP). Enhancement of immune activity in vitro and the adjuvanticity when inoculated with OVA were evaluated. The results demonstrated that the average molecular weight of AHP was 46.8 kDa and possessed typical polysaccharide absorption peaks. The entrapment efficiency for AHP within AHPP was 65.76 ± 3.31%. AHPP significantly stimulated phagocytic activity, MHCII and CD86 expression in macrophages. Further investigation showed that AHPP/OVA significantly enhanced lymphocyte proliferation and improved the CD4+/CD8+ T cell ratio. Moreover, AHPP/OVA treatment significantly increased IgG levels and up-regulated Th-associated cytokines with overall Th1 polarization. These studies demonstrated that AHP encapsulated within PLGA as a vaccine delivery system enhanced adaptive immunity.

Uji Akivitas Imunomodulator Ekstrak Buah Petai (Parkia speciosa Hassk.) terhadap Titer Imunoglobulin (IgG) pada Mencit Balb/c yang Diinduksi dengan SDMD

Immunomodulators are substances or drugs that can modulate the function and activity of the immune system. Most of the medicinal plants that have been studied prove the presence of immunostimulators, while immunosuppressors are still rare. Petai plants can function as antioxidants. There is a close relationship between antioxidants and the immune system giving antioxidants can stimulate and protect the cellular immune system. The purpose of this study was to determine the effect of administration and effective dose of petai fruit (Parkia speciosa Hassk) ethanol extract on increasing IgG titer in Balb / c mice induced by SDMD. This research method was started with Petai extraction and then tested for chemical content. Experimental animals consisted of 25 mice divided into 5 treatment groups, each group consisting of 5 mice. All groups were immunized with 1% SDMD as much as 0.2 ml / 20 g BB on day 8 (after being acclimatized for 7 days) intravenously. Test preparation is given orally for 7 days. On day 14, blood collection from retro orbital plexus was performed to collect serum and examined immunoglobulin G titers with indirect ELISA method. The benefit to be obtained from this study is to prove the ethanol extract of petai fruit (Parkia speciosa Hassk) can increase the immune system against foreign compounds with an increase in IgG titer in Balb / c mice induced by HRD. The results of Petai Fruit Extract Dosage 4.125 mg / KgBB, 8.25 mg / Kg BB and a dose of 16.5 mg / Kg BB give an effect on increasing IgG titers. The dose of 16.5 mg is better than stimouno in increasing IgG levels of mice induced by HRD. Keywords — Petai (Parkia speciosa Hassk), IgG, SDMD,ELISA

Accelerated Blood Clearance of Antibodies by Nanosized Click Antidotes.

Long blood half-life is one of the advantages of antibodies over small molecule drugs. At the same time, prolonged half-life is a problem for imaging applications or in the case of antibody-induced toxicities. There is a substantial need for antidotes that can quickly clear antibodies from systemic circulation and peripheral tissues. Engineered nanoparticles exhibit intrinsic affinity for clearance organs (mainly liver and spleen). trans-Cyclooctene (TCO) and methyltetrazine (MTZ) are versatile copper-free click chemistry components that are extensively being used for in vivo bioorthogonal couplings. To test the ability of nanoparticles to eliminate antibodies, we prepared a set of click-modified, clinically relevant antidotes based on several classes of drug carriers: phospholipid-PEG micelles, bovine serum albumin (BSA), and cross-linked dextran iron oxide (CLIO) nanoparticles. Mice were injected with IRDye 800CW-labeled, click-modified IgG followed by a click-modified antidote or PBS (control), and the levels of the IgG were monitored up to 72 h postinjection. Long-circulating lipid micelles produced a spike in IgG levels at 1 h, decreased IgG levels at 24 h, and did not decrease the area under the curve (AUC) and IgG accumulation in main organs. Long-circulating BSA decreased IgG levels at 1 and 24 h, decreased the AUC, but did not significantly decrease organ accumulation. Long-circulating CLIO nanoworms increased IgG levels at 1 h, decreased IgG levels at 24 h, did not decrease the AUC, and did not decrease the organ accumulation. On the other hand, short-circulating CLIO nanoparticles decreased IgG levels at 1 and 24 h, significantly decreasing the AUC and accumulation in the main organs. Multiple doses of CLIO and BSA were not able to completely eliminate the antibody from blood, despite the click reactivity of the residual IgG, likely due to exchange of IgG between blood and tissue compartments. Pharmacokinetic modeling suggests that short antidote half-life and fast click reaction rate should result in higher IgG depletion efficiency. Short-circulating click-modified nanocarriers are the most effective antidotes for elimination of antibodies from blood. This study sets a stage for future development of antidotes based on nanomedicine.

Immune responses elicited by the recombinant Erp, HspR, LppX, MmaA4, and OmpA proteins from Mycobacterium tuberculosis in mice.

Immunogenic potency of the recombinant Erp, HspR, LppX, MmaA4, and OmpA proteins from Mycobacterium tuberculosis (MTB), formulated with Montanide ISA 720 VG adjuvant, was evaluated in BALB/c mice for the first time in this study. The five vaccine formulations, adjuvant, and BCG vaccine were subcutaneously injected into mice, and the sera were collected at days 0, 15, 30, 41, and 66. The humoral and cellular immune responses against vaccine formulations were determined by measuring serum IgG and serum interferon-gamma (IFN-γ) and interleukin-12 (IL-12) levels, respectively. All formulations significantly increased IgG levels post-vaccination. The highest increase in IFN-γ level was provided by MmaA4 formulation. The Erp, HspR, and LppX formulations were as effective as BCG in enhancement of IFN-γ level. The most efficient vaccine boosting the IL-12 level was HspR formulation, especially at day 66. Erp formulation also increased the IL-12 level more than BCG at days 15 and 30. The IL-12 level boosted by MmaA4 formulation was found to be similar to that by BCG. OmpA formulation was inefficient in enhancement of cellular immune responses. This study showed that MmaA4, HspR, and Erp proteins from MTB are successful in eliciting both humoral and cellular immune responses in mice.

BK polyomavirus viremia and antibody responses of pediatric kidney transplant recipients in Finland.

BKPyV is an important cause of premature graft failure after KT. Most clinical studies describe BKPyV infection in adult KT patients. We studied the prevalence of post-transplant BKPyV viremia, serology, and graft function in pediatric KT recipients. Forty-six pediatric patients transplanted between 2009 and 2014 were followed up for BKPyV DNAemia by plasma PCR for median 2.3 (range: 1-6) years. BKPyV-specific antibodies were retrospectively analyzed using virus-like particle ELISA. GFR was measured annually by 51 Cr-EDTA clearance, and serum samples were screened for DSAs by Luminex assay. BKPyV viremia was demonstrated in nine patients at a median of 6months post-KT. Early BKPyV viremia at 3months post-KT associated with decreased concomitant GFR and tendency for decreased subsequent graft function. Three of nine patients with BKPyV viremia developed DSA, all against class II antigens. PyVAN developed to four patients and responded to judicious reduction in IS. One graft was lost later due to ABMR. BKPyV-IgG was found in 18 of 31 patients (58%) tested at transplantation, and seven recipients seroconverted after transplantation with a significant increase in IgG levels with IgM. Finally, BKPyV-IgG was detectable in 31 of 40 patients (78%) at the end of the study. Post-transplant BKPyV viremia in pediatric KT patients may alter graft function and contribute to progression of chronic allograft injury. BKPyV-IgG predicts past exposure. Low or absent BKPyV-specific antibody levels were seen pretransplant in 42% of tested patients, but were not predictive of prolonged replication or poor outcome.

Potential pitfalls in analyzing structural uncoupling of eNOS: aging is not associated with increased enzyme monomerization.

Homodimer formation is essential for the normal activity of endothelial nitric oxide synthase (eNOS). Structural uncoupling of eNOS, with generation of enzyme monomers, is thought to contribute to endothelial dysfunction in several vascular disorders, including aging. However, low-temperature SDS-PAGE of healthy arteries has revealed considerable variation between studies in the relative expression of eNOS dimers and monomers. While assessing structural uncoupling of eNOS in aging arteries, we identified methodological pitfalls that might contribute to such variation. Therefore, using human cultured aortic endothelial cells and aortas from young and aged Fischer-344 rats, we investigated optimal approaches for analyzing the expression of eNOS monomers and dimers. The results demonstrated that published differences in treatment of cell lysates can significantly impact the relative expression of several eNOS species, including denatured monomers, partially folded monomers, dimers, and higher-order oligomers. In aortas, experiments initially confirmed a large increase in eNOS monomers in aging arteries, consistent with structural uncoupling. However, these monomers were actually endogenous IgG, which, under these conditions, has mobility similar to eNOS monomers. Increased IgG levels in aged aortas likely reflect the aging-induced disruption of endothelial junctions and increased arterial penetration of IgG. After removal of the IgG signal, there were low levels of eNOS monomers in young arteries, which were not significantly different in aged arteries. Therefore, structural uncoupling of eNOS is not a prominent feature in young healthy arteries, and the process is not increased by aging. The study also identifies optimal approaches to analyze eNOS dimers and monomers. NEW & NOTEWORTHY Structural uncoupling of endothelial nitric oxide synthase (eNOS) is considered central to endothelial dysfunction. However, reported levels of eNOS dimers and monomers vary widely, even in healthy arteries. We demonstrate that sample processing can alter relative levels of eNOS species. Moreover, endothelial dysfunction in aging aortas results in IgG accumulation, which, because of similar mobility to eNOS monomers, could be misinterpreted as structural uncoupling. Indeed, enzyme monomerization is not prominent in young or aging arteries.

IgM and IgG against Plasmodium falciparum lysate as surrogates of malaria exposure and protection during pregnancy

BackgroundDifficulties to disentangle the protective versus exposure role of anti-malarial antibodies hamper the identification of clinically-relevant immune targets. Here, factors affecting maternal IgG and IgMs against Plasmodium falciparum antigens, as well as their relationship with parasite infection and clinical outcomes, were assessed in mothers and their children. Antibody responses among 207 Mozambican pregnant women at delivery against MSP119, EBA175, AMA1, DBLα and parasite lysate (3D7, R29 and E8B parasite lines), as well as the surface of infected erythrocytes, were assessed by enzyme-linked immunosorbent assay and flow cytometry. The relationship between antibody levels, maternal infection and clinical outcomes was assessed by multivariate regression analysis.ResultsPlacental infection was associated with an increase in maternal levels of IgGs and IgMs against a broad range of parasite antigens. The multivariate analysis including IgGs and IgMs showed that the newborn weight increased with increasing IgG levels against a parasite lysate, whereas the opposite association was found with IgMs. IgGs are markers of protection against poor pregnancy outcomes and IgMs of parasite exposure.ConclusionsAdjusting the analysis for the simultaneous effect of IgMs and IgGs can contribute to account for heterogeneous exposure to P. falciparum when assessing immune responses effective against malaria in pregnancy.

Characterization of the immune response elicited by the vaccinia virus L3 protein delivered as naked DNA

Poxviruses are complex dsDNA viruses with over 200 genes, many of them with unknown role in the stimulation of immune responses. Among these, the vaccinia virus (VACV) L3L ORF encodes an essential protein for the transcription of the VACV early genes. To the best of our knowledge, the immune response elicited by L3 has not been characterized. In this regard, our data describes a DNA L3-coding plasmid (pL3L) that stimulates both, humoral- and cell-mediated immune responses in a mouse model. Cell-mediated immune responses were measured by IFN-γ and IL-4 ELISPOT assays. We performed CD8+ cells depletion and flow cytometry analysis to account for the contribution of cytotoxic T lymphocytes in the IFN-γ production. Moreover, results from ELISPOT were confirmed by measuring the concentration of IL-4 and IFN-γ in supernatant of antigen-stimulated splenocytes by cytokine ELISA. Additionally, dominant antigenic regions of L3 protein were identified by epitope mapping analysis. Humoral immune responses were assessed by ELISA. Specifically, the production of total IgG, IgG1 (TH-2) and IgG2a (TH-1) were determined one week after the final immunization. Our ELISPOT data shows pL3L-immunized animals to produce significantly higher frequencies of IFN-γ Spot-Forming Cells (SFC) versus controls. IL-4 levels remained unchanged in all three groups, demonstrating the increase in antigen-specific IFN-γ releasing cells. Flow cytometry assay results showed that CD8+ T cells are a major contributor to the production of IFN-γ. Moreover, our formulation enhances the production of total IgG, predominantly IgG2a isotype. Immunization with pL3L promotes a robust cytotoxic immune response, crucial against viral pathogens. In addition, our vaccine candidate promotes an increase in IgG levels, especially IgG2a (TH-1 type). Our data encourages further studies of L3 as a novel antigen in vaccine development against poxviruses.