The key factor in the development of antibody-based assays is to find an antibody that has an appropriate affinity, high specificity, and low cross-reactivity. However, this task is not easy to carry out since the research antibodies on the market may suffer from low specificity and reproducibility. Here, we report on a palm-sized dot blot-based device, called the affiblot, that has a specially designed lid that allows simultaneous semi-quantitative comparison of up to five antibodies from different suppliers regarding their affinity/avidity, cross-reactivity, and batch-to-batch reliability. The only required peripheral equipment is a vacuum pump, a camera, and densitometry software. The affiblot device was tested for its functionality and its measurements were compared against those obtained by standard dot blot and ELISA. The benefit over these methods, when various antibodies are evaluated, is in its simplicity. It allows easy antigen deposition, fast application and the discarding of the solutions, a compact undivided membrane, and therefore significant decrease of labor. The device was tested with specific anti-ApoE, anti-EpCAM, anti-Salmonella, anti-E. coli, and anti-Listeria antibodies from different suppliers. Their properties were compared for their ability to interact specifically with antigen and/or non-target structures and the best-suited antibody for the intended application was identified.
- MeSH
- ELISA MeSH
- imunoblotting MeSH
- monoklonální protilátky * MeSH
- reprodukovatelnost výsledků MeSH
- zkřížené reakce MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- monoklonální protilátky * MeSH
The adaptive immune response to severe acute respiratory coronavirus 2 (SARS-CoV-2) is important for vaccine development and in the recovery from coronavirus disease 2019 (COVID-19). Men and cancer patients have been reported to be at higher risks of contracting the virus and developing the more severe forms of COVID-19. Prostate cancer (PCa) may be associated with both of these risks. We show that CD4+ T cells of SARS-CoV-2-unexposed patients with hormone-refractory (HR) metastatic PCa had decreased CD4+ T cell immune responses to antigens from SARS-CoV-2 spike glycoprotein but not from the spiked glycoprotein of the 'common cold'-associated human coronavirus 229E (HCoV-229E) as compared with healthy male volunteers who responded comparably to both HCoV-229E- and SARS-CoV-2-derived antigens. Moreover, the HCoV-229E spike glycoprotein antigen-elicited CD4+ T cell immune responses cross-reacted with the SARS-CoV-2 spiked glycoprotein antigens. PCa patients may have impaired responses to the vaccination, and the cross-reactivity can mediate antibody-dependent enhancement (ADE) of COVID-19. These findings highlight the potential for increased vulnerability of PCa patients to COVID-19.
- Klíčová slova
- COVID-19, HCoV-229E, SARS-CoV-2, prostate cancer, spike glycoprotein,
- MeSH
- adaptivní imunita MeSH
- CD4-pozitivní T-lymfocyty imunologie MeSH
- CD8-pozitivní T-lymfocyty imunologie MeSH
- COVID-19 imunologie virologie MeSH
- cytokiny imunologie MeSH
- glykoprotein S, koronavirus imunologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- lidský koronavirus 229E imunologie MeSH
- nádory prostaty imunologie patologie MeSH
- SARS-CoV-2 imunologie MeSH
- senioři MeSH
- zkřížené reakce MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- cytokiny MeSH
- glykoprotein S, koronavirus MeSH
- spike protein, SARS-CoV-2 MeSH Prohlížeč
Tick-borne encephalitis virus (TBEV) is an emerging human pathogen that causes potentially fatal disease with no specific treatment. Mouse monoclonal antibodies are protective against TBEV, but little is known about the human antibody response to infection. Here, we report on the human neutralizing antibody response to TBEV in a cohort of infected and vaccinated individuals. Expanded clones of memory B cells expressed closely related anti-envelope domain III (EDIII) antibodies in both groups of volunteers. However, the most potent neutralizing antibodies, with IC50s below 1 ng/ml, were found only in individuals who recovered from natural infection. These antibodies also neutralized other tick-borne flaviviruses, including Langat, louping ill, Omsk hemorrhagic fever, Kyasanur forest disease, and Powassan viruses. Structural analysis revealed a conserved epitope near the lateral ridge of EDIII adjoining the EDI-EDIII hinge region. Prophylactic or early therapeutic antibody administration was effective at low doses in mice that were lethally infected with TBEV.
- MeSH
- analýza přežití MeSH
- epitopy imunologie MeSH
- imunoglobulin G aplikace a dávkování imunologie MeSH
- klíšťová encefalitida imunologie prevence a kontrola virologie MeSH
- kohortové studie MeSH
- kultivované buňky MeSH
- lidé MeSH
- monoklonální protilátky aplikace a dávkování genetika imunologie MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- neutralizující protilátky aplikace a dávkování genetika imunologie MeSH
- proteiny virového obalu genetika imunologie MeSH
- protilátky virové aplikace a dávkování genetika imunologie MeSH
- sekvence aminokyselin MeSH
- sekvenční homologie aminokyselin MeSH
- viry klíšťové encefalitidy účinky léků imunologie fyziologie MeSH
- zkřížené reakce imunologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
- Názvy látek
- epitopy MeSH
- imunoglobulin G MeSH
- monoklonální protilátky MeSH
- neutralizující protilátky MeSH
- proteiny virového obalu MeSH
- protilátky virové MeSH
COVID-19 is a global pandemic caused by the SARS-CoV-2 coronavirus. T cells play a key role in the adaptive antiviral immune response by killing infected cells and facilitating the selection of virus-specific antibodies. However, neither the dynamics and cross-reactivity of the SARS-CoV-2-specific T-cell response nor the diversity of resulting immune memory is well understood. In this study, we use longitudinal high-throughput T-cell receptor (TCR) sequencing to track changes in the T-cell repertoire following two mild cases of COVID-19. In both donors, we identified CD4+ and CD8+ T-cell clones with transient clonal expansion after infection. We describe characteristic motifs in TCR sequences of COVID-19-reactive clones and show preferential occurrence of these motifs in publicly available large dataset of repertoires from COVID-19 patients. We show that in both donors, the majority of infection-reactive clonotypes acquire memory phenotypes. Certain T-cell clones were detected in the memory fraction at the pre-infection time point, suggesting participation of pre-existing cross-reactive memory T cells in the immune response to SARS-CoV-2.
- Klíčová slova
- COVID-19, RepSeq, SARS-CoV-2, TCR, computational biology, human, immunology, inflammation, systems biology,
- MeSH
- COVID-19 imunologie patofyziologie MeSH
- genová knihovna MeSH
- imunologická paměť * MeSH
- lidé MeSH
- longitudinální studie MeSH
- mapování epitopu MeSH
- receptory antigenů T-buněk chemie genetika MeSH
- SARS-CoV-2 fyziologie MeSH
- sekvence aminokyselin MeSH
- stupeň závažnosti nemoci MeSH
- T-lymfocyty imunologie MeSH
- testování histokompatibility MeSH
- zkřížené reakce MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- receptory antigenů T-buněk MeSH
BACKGROUND: Walnut allergy is common across the globe, but data on the involvement of individual walnut components are scarce. OBJECTIVES: To identify geographical differences in walnut component sensitization across Europe, explore cosensitization and cross-reactivity, and assess associations of clinical and serological determinants with severity of walnut allergy. METHODS: As part of the EuroPrevall outpatient surveys in 12 European cities, standardized clinical evaluation was conducted in 531 individuals reporting symptoms to walnut, with sensitization to all known walnut components assessed in 202 subjects. Multivariable Lasso regression was applied to investigate predictors for walnut allergy severity. RESULTS: Birch-pollen-related walnut sensitization (Jug r 5) dominated in Northern and Central Europe and lipid transfer protein sensitization (Jug r 3) in Southern Europe. Profilin sensitization (Jug r 7) was prominent throughout Europe. Sensitization to storage proteins (Jug r 1, 2, 4, and 6) was detected in up to 10% of subjects. The walnut components that showed strong correlations with pollen and other foods differed between centers. The combination of determinants best predicting walnut allergy severity were symptoms upon skin contact with walnut, atopic dermatitis (ever), family history of atopic disease, mugwort pollen allergy, sensitization to cat or dog, positive skin prick test result to walnut, and IgE to Jug r 1, 5, 7, or carbohydrate determinants (area under the curve = 0.81; 95% CI, 0.73-0.89). CONCLUSIONS: Walnut-allergic subjects across Europe show clear geographical differences in walnut component sensitization and cosensitization patterns. A predictive model combining results from component-based serology testing with results from extract-based testing and information on clinical background allows for good discrimination between mild to moderate and severe walnut allergy.
- Klíčová slova
- Allergen components, EuroPrevall, Europe, IgE sensitization, Prediction, Severity, Walnut allergy, iFAAM,
- MeSH
- alergeny MeSH
- antigeny rostlinné MeSH
- imunoglobulin E MeSH
- Juglans * MeSH
- kočky MeSH
- lidé MeSH
- ořechy * MeSH
- potravinová alergie * MeSH
- psi MeSH
- zkřížené reakce MeSH
- zvířata MeSH
- Check Tag
- kočky MeSH
- lidé MeSH
- psi MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Evropa epidemiologie MeSH
- Názvy látek
- alergeny MeSH
- antigeny rostlinné MeSH
- imunoglobulin E MeSH
The discovery of T-cell responses to SARS-CoV-2 in non-infected individuals indicates cross-reactive immune memory from prior exposure to human coronaviruses (HCoV) that cause the common cold. This raises the possibility that "immunity" could exist within populations at rates that may be higher than serology studies estimate. Besides specialized research labs, however, there is limited ability to measure HCoV CD4+ and CD8+ T-cell responses to SARS-CoV-2 infection, which currently impedes interpretation of any potential correlation between COVID-19 disease pathogenesis and the calibration of pandemic control measures. Given this limited testing ability, an alternative approach would be to exploit the large cohort of currently available data from which statistically significant associations may be generated. This would necessitate the merging of several public databases including patient and contact tracing, which could be created by relevant public health organizations. Including data from both symptomatic and asymptomatic patients in SARS-CoV-2 databases and surveillance systems could provide the necessary information to allow for more informed decisions.
- Klíčová slova
- COVID-19, SARS-CoV-2, T-cell, children, contact tracing, human coronaviruses, immunity,
- MeSH
- COVID-19 imunologie virologie MeSH
- kohortové studie MeSH
- lidé MeSH
- nachlazení imunologie MeSH
- SARS-CoV-2 fyziologie MeSH
- T-lymfocyty imunologie MeSH
- zkřížené reakce MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Recent studies have shown a significant level of T cell immunity to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in convalescent coronavirus disease 2019 (COVID-19) patients and unexposed healthy individuals. Also, SARS-CoV-2-reactive T memory cells occur in unexposed healthy individuals from endemic coronaviruses that cause the 'common cold.' The finding of the expression of adaptive SARS-CoV-2-reactive T memory cells in unexposed healthy individuals may be due to multiple cross-reactive viral protein targets following previous exposure to endemic human coronavirus infections. The opinion of the authors is that determination of protein sequence homologies across seemingly disparate viral protein libraries may provide epitope-matching data that link SARS-CoV-2-reactive T memory cell signatures to prior administration of cross-reacting vaccines to common viral pathogens. Exposure to SARS-CoV-2 initiates diverse cellular immune responses, including the associated 'cytokine storm'. Therefore, it is possible that the intact virus possesses a required degree of conformational matching, or stereoselectivity, to effectively target its receptor on multiple cell types. Therefore, conformational matching may be viewed as an evolving mechanism of viral infection and viral replication by an evolutionary modification of the angiotensin-converting enzyme 2 (ACE2) receptor required for SARS-CoV-2 binding and host cell entry. The authors propose that convalescent memory T cell immunity in individuals with mild or asymptomatic SARS-CoV-2 infection may result from an evolutionarily adapted immune response to coronavirus and the 'common cold'.
- MeSH
- angiotensin konvertující enzym 2 genetika MeSH
- asymptomatické infekce * MeSH
- buněčná imunita genetika MeSH
- COVID-19 krev diagnóza imunologie virologie MeSH
- epitopy T-lymfocytární genetika imunologie MeSH
- imunogenicita vakcíny MeSH
- imunologická paměť genetika MeSH
- internalizace viru MeSH
- lidé MeSH
- molekulární evoluce MeSH
- nachlazení imunologie prevence a kontrola virologie MeSH
- protilátky virové MeSH
- replikace viru genetika imunologie MeSH
- Rhinovirus genetika imunologie MeSH
- SARS-CoV-2 imunologie patogenita MeSH
- sekvenční homologie MeSH
- stupeň závažnosti nemoci MeSH
- T-lymfocyty - podskupiny imunologie MeSH
- T-lymfocyty imunologie MeSH
- virové proteiny genetika imunologie MeSH
- virové vakcíny aplikace a dávkování imunologie MeSH
- zkřížené reakce genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- ACE2 protein, human MeSH Prohlížeč
- angiotensin konvertující enzym 2 MeSH
- epitopy T-lymfocytární MeSH
- protilátky virové MeSH
- virové proteiny MeSH
- virové vakcíny MeSH
INTRODUCTION: Frequently observed multiple sensitizations to several animals highlights the importance of a molecular diagnosis, distinguishing between sensitizations specific to single species and sensitizations due to cross-reactivity. OBJECTIVE: The aim of our study was to assess the usefulness of a molecular diagnosis in the description of sensitization profiles in allergy patients living in Central Europe, with a particular focus on animal-derived molecules. METHODS: The molecular diagnosis was performed using the ImmunoCAP ISAC microarray. Results of 1,255 allergy patients were subjected to statistical analysis. RESULTS: The highest sensitization rates were observed for uteroglobin Fel d 1 (31.8%) and kallikrein Can f 5 (16.4%), followed by animal lipocalins Can f 1 (13.9%), Equ c 1 (6.2%), Fel d 4 (5.3%), Can f 2 (4.2%), and Mus m 1 (4.1%). Sensitization rates to serum albumins Fel d 2, Can f 3, Equ c 3, and Bos d 6 were very low, with the highest being 3.2% to Fel d 2. Detailed subanalysis confirmed the dominant role of Fel d 1 or Can f 5 and/or Can f 1 in cat- or dog-sensitized patients, respectively. Further analysis focused on lipocalins and albumins confirmed a high rate of cosensitizations within both groups. CONCLUSION: The sensitization to animal allergen molecules is very frequent in Central Europe. The most common is sensitization to species-specific cat uteroglobin Fel d 1 and dog kallikrein Can f 5, followed by sensitizations to animal lipocalins. Our data suggest that commonly observed multiple sensitizations detected by extract approach can be explained not only by true cosensitization, but also by cross-reactivity, mainly in the frame of lipocalins. Cross-reactive serum albumins are minor sensitizers and are probably not important from this point of view.
- Klíčová slova
- Animal allergen, Animal allergy, Molecular diagnostics, Sensitization profile, Specific IgE,
- MeSH
- alergeny imunologie MeSH
- alergie imunologie MeSH
- dítě MeSH
- dospělí MeSH
- druhová specificita MeSH
- kočky MeSH
- kojenec MeSH
- lidé středního věku MeSH
- lidé MeSH
- lipokaliny imunologie MeSH
- mladiství MeSH
- mladý dospělý MeSH
- myši MeSH
- předškolní dítě MeSH
- psi MeSH
- senioři MeSH
- sérový albumin imunologie MeSH
- zkřížené reakce imunologie MeSH
- zvířata MeSH
- Check Tag
- dítě MeSH
- dospělí MeSH
- kočky MeSH
- kojenec MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- myši MeSH
- předškolní dítě MeSH
- psi MeSH
- senioři MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Evropa MeSH
- Názvy látek
- alergeny MeSH
- lipokaliny MeSH
- sérový albumin MeSH
Birch and other related trees of the families Betulaceae and Fagaceae (alder, hazel, oak, hornbeam, chestnut, and beech) constitute the birch homologous group. This grouping is primarily based on the extensive IgE cross-reactivity of allergen homologs to the major birch allergen Bet v 1. Birch pollen is the most dominant tree pollen in Northern and Central Europe and is a major cause of allergic rhinitis and, possibly, asthma symptoms. Over the last few decades, levels of birch pollen have risen and the period of exposure has increased due to climate changes. Subsequently, the prevalence of birch pollen sensitization has also increased. The cross-reactivity and sequential pollen seasons within the birch homologous group create a prolonged symptomatic allergy period beyond birch pollen alone. Furthermore, many plant food allergens contain homologs to Bet v 1, meaning that the majority of patients with birch pollen allergy suffer from secondary pollen food syndrome (PFS). As a result, the negative impact on health-related quality of life (HRQoL) in patients allergic to birch pollen is significant. The purpose of this manuscript was to narratively review topics of interest such as taxonomy, cross-reactivity, prevalence, clinical relevance, PFS, and HRQoL with regard to birch pollen allergy from a European perspective.
- Klíčová slova
- alder, allergic rhinitis, birch, cross-reactivity, hazel,
- MeSH
- alergeny imunologie MeSH
- antigeny rostlinné imunologie MeSH
- bříza imunologie MeSH
- imunizace MeSH
- imunoglobulin E imunologie MeSH
- lidé MeSH
- ochrana veřejného zdraví MeSH
- prevalence MeSH
- pyl imunologie MeSH
- roční období MeSH
- sezónní alergická rýma diagnóza epidemiologie imunologie MeSH
- určení symptomu MeSH
- zkřížené reakce imunologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Geografické názvy
- Evropa epidemiologie MeSH
- Názvy látek
- alergeny MeSH
- antigeny rostlinné MeSH
- imunoglobulin E MeSH
The α-Gal syndrome (AGS) is a type of allergy characterized by an IgE antibody (Ab) response against the carbohydrate Galα1-3Galβ1-4GlcNAc-R (α-Gal), which is present in glycoproteins from tick saliva and tissues of non-catarrhine mammals. Recurrent tick bites induce high levels of anti-α-Gal IgE Abs that mediate delayed hypersensitivity to consumed red meat products in humans. This was the first evidence that tick glycoproteins play a major role in allergy development with the potential to cause fatal delayed anaphylaxis to α-Gal-containing foods and drugs and immediate anaphylaxis to tick bites. Initially, it was thought that the origin of tick-derived α-Gal was either residual blood meal mammalian glycoproteins containing α-Gal or tick gut bacteria producing this glycan. However, recently tick galactosyltransferases were shown to be involved in α-Gal synthesis with a role in tick and tick-borne pathogen life cycles. The tick-borne pathogen Anaplasma phagocytophilum increases the level of tick α-Gal, which potentially increases the risk of developing AGS after a bite by a pathogen-infected tick. Two mechanisms might explain the production of anti-α-Gal IgE Abs after tick bites. The first mechanism proposes that the α-Gal antigen on tick salivary proteins is presented to antigen-presenting cells and B-lymphocytes in the context of Th2 cell-mediated immunity induced by tick saliva. The second mechanism is based on the possibility that tick salivary prostaglandin E2 triggers Immunoglobulin class switching to anti-α-Gal IgE-producing B cells from preexisting mature B cells clones producing anti-α-Gal IgM and/or IgG. Importantly, blood group antigens influence the capacity of the immune system to produce anti-α-Gal Abs which in turn impacts individual susceptibility to AGS. The presence of blood type B reduces the capacity of the immune system to produce anti-α-Gal Abs, presumably due to tolerance to α-Gal, which is very similar in structure to blood group B antigen. Therefore, individuals with blood group B and reduced levels of anti-α-Gal Abs have lower risk to develop AGS. Specific immunity to tick α-Gal is linked to host immunity to tick bites. Basophil activation and release of histamine have been implicated in IgE-mediated acquired protective immunity to tick infestations and chronic itch. Basophil reactivity was also found to be higher in patients with AGS when compared to asymptomatic α-Gal sensitized individuals. In addition, host resistance to tick infestation is associated with resistance to tick-borne pathogen infection. Anti-α-Gal IgM and IgG Abs protect humans against vector-borne pathogens and blood group B individuals seem to be more susceptible to vector-borne diseases. The link between blood groups and anti-α-Gal immunity which in turn affects resistance to vector-borne pathogens and susceptibility to AGS, suggests a trade-off between susceptibility to AGS and protection to some infectious diseases. The understanding of the environmental and molecular drivers of the immune mechanisms involved in AGS is essential to developing tools for the diagnosis, control, and prevention of this growing health problem.
- Klíčová slova
- IgE, food allergy, red meat allergy, ticks, α-Gal syndrome (AGS),
- MeSH
- alergeny imunologie MeSH
- anafylaxe etiologie MeSH
- červené maso MeSH
- genetická predispozice k nemoci MeSH
- hmyzí proteiny imunologie MeSH
- imunoglobulin E metabolismus MeSH
- interakce genů a prostředí MeSH
- klíšťata MeSH
- kousnutí klíštětem komplikace imunologie MeSH
- lidé MeSH
- potravinová alergie komplikace etiologie imunologie MeSH
- tvorba protilátek MeSH
- zkřížené reakce MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Názvy látek
- alergeny MeSH
- hmyzí proteiny MeSH
- imunoglobulin E MeSH