A subgroup of coeliac disease patients continues to experience symptoms even on a gluten-free diet (GFD). We attempted to determine whether these symptoms could be due to either cross-contamination with gluten-contain...A subgroup of coeliac disease patients continues to experience symptoms even on a gluten-free diet (GFD). We attempted to determine whether these symptoms could be due to either cross-contamination with gluten-containing foods or cross-reactivity between α-gliadin and non-gluten foods consumed on a GFD. We measured the reactivity of affinity-purified polyclonal and monoclonal α-gliadin 33-mer peptide antibodies against gliadin and additional food antigens commonly consumed by patients on a GFD using ELISA and dot-blot. We also examined the immune reactivity of these antibodies with various tissue antigens. We observed significant immune reactivity when these antibodies were applied to cow’s milk, milk chocolate, milk butyrophilin, whey protein, casein, yeast, oats, corn, millet, instant coffee and rice. To investigate whether there was cross-reactivity between α-gliadin antibody and different tissue antigens, we measured the degree to which this antibody bound to these antigens. The most significant binding occurred with asialoganglioside, hepatocyte, glutamic acid decarboxylase 65, adrenal 21-hydroxylase, and various neural antigens. The specificity of anti-α-gliadin binding to different food and tissue antigens was demonstrated by absorption and inhibition studies. We also observed significant cross-reactivity between α-gliadin 33-mer and various food antigens, but some of these reactions were associated with the contamination of non-gluten foods with traces of gluten. The consumption of cross-reactive foods as well as gluten-contaminated foods may be responsible for the continuing symptoms presented by a subgroup of patients with coeliac disease. The lack of response of some CD patients may also be due to antibody cross-reactivity with non-gliadin foods. These should then be treated as gluten-like peptides and should also be excluded from the diet when the GFD seems to fail.展开更多
To investigate the significance of the SARS-associated coronavirus (SARS-CoV) antibody,detected by ELISA and indirect immunofluorescence assays (IFA) for the SARS-CoV Vero E6 cell lysates,in non-SARS subjects, 114 ser...To investigate the significance of the SARS-associated coronavirus (SARS-CoV) antibody,detected by ELISA and indirect immunofluorescence assays (IFA) for the SARS-CoV Vero E6 cell lysates,in non-SARS subjects, 114 serum samples from healthy controls and 104 serum specimens from autoimmune disease patients were collected.The results of ELISA showed that among 114 sera from healthy controls,4 (3.5%) were positive of SARS-CoV-IgG antibody and 114 (100%) were all negative of SARS-CoV-IgM antibody;the specificity of SARS-CoV-IgG antibody for SARS patients was 96.5%,but the specificity of both SARS-CoV-IgG and -IgM antibodies for SARS patients was 100%.In 58 cases with SLE,positive rates of SARS-CoV-IgG and -IgM antibodies were 32.8% (19/58) and 8.6% (5/58),respectively,in which 11 cases (19%) were positive of both SARS-CoV-IgG and -IgM antibodies;in 10 cases with SS,positive rate of both SARS-CoV-IgG and -IgM antibodies was 10% (1/10);in 16 cases with MCTD,positive rate of SARS-CoV-IgG was 37.5% (6/16),positive rate of both SARS-CoV-IgG and -IgM antibodies was 6.3% (1/16);in 20 cases with RA,one case was positive (5%) of SARS-CoV-IgG.However,of all samples with positive SARS-CoV-IgG and -IgM antibodies for autoimmune diseases and healthy controls,SARS-CoV RNA and antibodies were all negative by RT-PCR and IFA.All sera for negative or positive ELISA results were also negative or positive results using ELISA with Vero E6 cells lysates.These studies showed that SARS-CoV Vero E6 cell lysates for the ELISA to detect SARS-CoV antibodies could lead to the false-positive reactions or cross-reactions of SARS-CoV antibodies in non-SARS diseases and healthy controls,and the false-positive reactions or cross-reactions were related to Vero E6 cell lysates and autoantibodies in non-SARS population.Cellular & Molecular Immunology.2004;1(4): 304-307.展开更多
This review aimed to assess the occurrence of false-positive serological reaction between dengue and coronavirus disease 2019(COVID-19)and its implications for diagnosis.Evidence syntheses were conducted by systematic...This review aimed to assess the occurrence of false-positive serological reaction between dengue and coronavirus disease 2019(COVID-19)and its implications for diagnosis.Evidence syntheses were conducted by systematically reviewing available literature using multiple databases,including Web of Science,PubMed,Google Scholar and medRxiv.Among a total of 16 presented cases from clinical settings,cross-reaction to COVID-19 serological tests was observed in two(12.5%)dengue-positive patients,while 14 patients(87.5%)confirmed positive for severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)showed a cross-reaction with dengue serological tests,leading to misdiagnosis and mismanagement by attending clinicians.Of 1789 SARS-CoV-2-positive sera,cross-reaction to dengue serological tests was observed in 180 sera(10%),which is higher than the cross-reaction observed for SARS-CoV-2 in archived pre-COVID-19 sera positive for a dengue infection(75 of 811,9.2%,P=0.674).Clinicians in tropical regions are therefore advised to interpret serological tests with caution and use a more pragmatic approach to triage these infections.展开更多
In patients with respiratory allergy,cross-reactivity between aeroallergens and foods may induce food allergy,symptoms ranging from oral allergy syndrome to severe anaphylaxis.Clinical entities due to Ig E sensitizati...In patients with respiratory allergy,cross-reactivity between aeroallergens and foods may induce food allergy,symptoms ranging from oral allergy syndrome to severe anaphylaxis.Clinical entities due to Ig E sensitization to cross-reactive aeroallergen and food allergen components are described for many sources of plant origin(pollen-food syndromes and associations,such as birch-apple,cypress-peach and celery-mugwortspice syndromes,and mugwort-peach,mugwortchamomile,mugwort-mustard,ragweed-melon-banana,goosefoot-melon associations),fungal origin(Alternariaspinach syndrome),and invertebrate,mammalian or avian origin(mite-shrimp,cat-pork,and bird-egg syndromes).Clinical cases of allergic reactions to ingestion of food products containing pollen grains of specific plants,in patients with respiratory allergy to Asteraceae pollen,especially mugwort and ragweed,are also mentioned,for honey,royal jelly and bee polen dietary supplements,along with allergic reactions to foods contaminated with mites or fungi in patients with respiratory allergy to these aeroallergens.Medical history and diagnosis approach may be guided by the knowledge about the diverse cross-reacting allergens involved,and by the understanding of these clinical entities which may vary significantly or may be overlapping.The association between primary Ig E sensitization with respiratory symptoms to inhaled allergens and food allergy due to cross-reactive allergen components is important to assess in allergy practice.The use of molecular-based diagnosis improves the understanding of clinically relevant Ig E sensitization to cross-reactive allergen components from aeroallergen sources and foods.展开更多
AIM:To evaluate the presence and cross-reactive anti-bodies against hypervariable region 1(HVR1) in hepatitis C virus(HCV) infected patients and its relationship with the progression of the disease.METHODS:Sixteen rep...AIM:To evaluate the presence and cross-reactive anti-bodies against hypervariable region 1(HVR1) in hepatitis C virus(HCV) infected patients and its relationship with the progression of the disease.METHODS:Sixteen representative HVR1 proteins selected from a unique set of 1600 natural sequences were used to semiquantitate the cross-reactivity of HVR1 antibodies in the sera of HCV patients.Fifty-five chronic HCV patients including 23 with asymptomatic mild hepatitis,18 with chronic hepatitis and 16 with liver cirrhosis patients were studied.RESULTS:The degree of the cross-reactivity of anti-HVR1 antibodies in 23 patients with mild asymptomatic hepatitis was 3.09 ± 2.68,which was signif icantly lower than in those with chronic hepatitis(5.44 ± 3.93,P < 0.05) and liver cirrhosis(7.44 ± 3.90,P < 0.01).No correlation was observed between the broadness of the cross-reactivity anti-HVR1 antibodies and patient's age,infection time,serum alanine aminotransferase activity,or serum HCV-RNA concentration.It was the breath of cross-reactivity rather than the presence of anti-HVR1 antibody in HCV sera that was associated with the progression of liver disease.CONCLUSION:The broadly cross-reactive HVR1 antibodies generated in natural HCV patients can not neutralize the virus,which results in persistent infection in patients with chronic hepatitis.展开更多
An ester activation method was employed to couple enrofloxacin(ENFX) to the carrier proteins BSA and OVA. The conjugates ENFX-BSA and ENFX-OVA were identified with an UV spectrophotometer and amino acid automation a...An ester activation method was employed to couple enrofloxacin(ENFX) to the carrier proteins BSA and OVA. The conjugates ENFX-BSA and ENFX-OVA were identified with an UV spectrophotometer and amino acid automation analysis instrument, and resulted in conjugates with 48 ENFX molecules per carrier molecule(BSA). Splenocytes from mice immunized with ENFX-BSA were fused with SP2/0 myeloma cells and hybridomas secreting antibodies against enrofloxacin were selected and cloned. Two stable monoclonal antibodies, 2C5, 5D5 of the subclass IgG2a, were isolated. Using antibody 5D5, an indirect competitive inhibition enzyme-linked immunosorbent assay (Ci-ELISA) was developed for the quantitative detection of enrofloxacin and its metabolites. The IC50 of the standard curve was 21.67 ng mL-1 and the limit of detection for enrofloxacin was 0.13 ng mL-1. This method was sensitive and had a linear range from 0.13 to 10 000 ng mL-1 (r= -0.9782). Monoclonal antibody 5D5 exhibited high relative affinity to enrofloxacin, and the cross-reactivities with ciprofloxacin, marbofloxacin, sarafloxacin and danorfloxacin were 110.8, 27.40, 71.05 and 37.41%, respectively. Three non-fluoroquinolones of cefadroxil, chloramphenicol, sulfadimethoxine were tested and there was no cross-reaction between them.展开更多
Objectives To evaluate the immunogenicity of Mycobacterium intracellulare proteins and determine the cross-reactive proteins between M.intracellulare and M.tuberculosis.Methods Protein extracts from M.intracellulare w...Objectives To evaluate the immunogenicity of Mycobacterium intracellulare proteins and determine the cross-reactive proteins between M.intracellulare and M.tuberculosis.Methods Protein extracts from M.intracellulare were used to immunize BALB/c mice.The antigens were evaluated using cellular and humoral immunoassays.The common genes between M.intracellular and M.tuberculosis were identified using genome-wide comparative analysis,and cross-reactive proteins were screened using immunoproteome microarrays.Results Immunization with M.intracellulare proteins induced significantly higher levels of the cytokines interferon-γ(IFN-γ),interleukin-2(IL-2),interleukin-12(IL-12),interleukin-6(IL-6)and immunoglobulins IgG,IgG1,IgM,and IgG2a in mouse serum.Bone marrow-derived macrophages isolated from mice immunized with M.intracellulare antigens displayed significantly lower bacillary loads than those isolated from mice immunized with adjuvants.Whole-genome sequence analysis revealed 396 common genes between M.intracellulare and M.tuberculosis.Microchip hybridization with M.tuberculosis proteins revealed the presence of 478 proteins in the serum of mice immunized with M.intracellulare protein extracts.Sixty common antigens were found using both microchip and genomic comparative analyses.Conclusion This is the advanced study to investigate the immunogenicity of M.intracellulare proteins and the cross-reactive proteins between M.intracellulare and M.tuberculosis.The results revealed the presence of a number of cross-reactive proteins between M.intracellulare and M.tuberculosis.Therefore,this study provides a new way of identifying immunogenic proteins for use in tuberculosis vaccines against both M.intracellulare and M.tuberculosis in future.展开更多
文摘A subgroup of coeliac disease patients continues to experience symptoms even on a gluten-free diet (GFD). We attempted to determine whether these symptoms could be due to either cross-contamination with gluten-containing foods or cross-reactivity between α-gliadin and non-gluten foods consumed on a GFD. We measured the reactivity of affinity-purified polyclonal and monoclonal α-gliadin 33-mer peptide antibodies against gliadin and additional food antigens commonly consumed by patients on a GFD using ELISA and dot-blot. We also examined the immune reactivity of these antibodies with various tissue antigens. We observed significant immune reactivity when these antibodies were applied to cow’s milk, milk chocolate, milk butyrophilin, whey protein, casein, yeast, oats, corn, millet, instant coffee and rice. To investigate whether there was cross-reactivity between α-gliadin antibody and different tissue antigens, we measured the degree to which this antibody bound to these antigens. The most significant binding occurred with asialoganglioside, hepatocyte, glutamic acid decarboxylase 65, adrenal 21-hydroxylase, and various neural antigens. The specificity of anti-α-gliadin binding to different food and tissue antigens was demonstrated by absorption and inhibition studies. We also observed significant cross-reactivity between α-gliadin 33-mer and various food antigens, but some of these reactions were associated with the contamination of non-gluten foods with traces of gluten. The consumption of cross-reactive foods as well as gluten-contaminated foods may be responsible for the continuing symptoms presented by a subgroup of patients with coeliac disease. The lack of response of some CD patients may also be due to antibody cross-reactivity with non-gliadin foods. These should then be treated as gluten-like peptides and should also be excluded from the diet when the GFD seems to fail.
文摘To investigate the significance of the SARS-associated coronavirus (SARS-CoV) antibody,detected by ELISA and indirect immunofluorescence assays (IFA) for the SARS-CoV Vero E6 cell lysates,in non-SARS subjects, 114 serum samples from healthy controls and 104 serum specimens from autoimmune disease patients were collected.The results of ELISA showed that among 114 sera from healthy controls,4 (3.5%) were positive of SARS-CoV-IgG antibody and 114 (100%) were all negative of SARS-CoV-IgM antibody;the specificity of SARS-CoV-IgG antibody for SARS patients was 96.5%,but the specificity of both SARS-CoV-IgG and -IgM antibodies for SARS patients was 100%.In 58 cases with SLE,positive rates of SARS-CoV-IgG and -IgM antibodies were 32.8% (19/58) and 8.6% (5/58),respectively,in which 11 cases (19%) were positive of both SARS-CoV-IgG and -IgM antibodies;in 10 cases with SS,positive rate of both SARS-CoV-IgG and -IgM antibodies was 10% (1/10);in 16 cases with MCTD,positive rate of SARS-CoV-IgG was 37.5% (6/16),positive rate of both SARS-CoV-IgG and -IgM antibodies was 6.3% (1/16);in 20 cases with RA,one case was positive (5%) of SARS-CoV-IgG.However,of all samples with positive SARS-CoV-IgG and -IgM antibodies for autoimmune diseases and healthy controls,SARS-CoV RNA and antibodies were all negative by RT-PCR and IFA.All sera for negative or positive ELISA results were also negative or positive results using ELISA with Vero E6 cells lysates.These studies showed that SARS-CoV Vero E6 cell lysates for the ELISA to detect SARS-CoV antibodies could lead to the false-positive reactions or cross-reactions of SARS-CoV antibodies in non-SARS diseases and healthy controls,and the false-positive reactions or cross-reactions were related to Vero E6 cell lysates and autoantibodies in non-SARS population.Cellular & Molecular Immunology.2004;1(4): 304-307.
文摘This review aimed to assess the occurrence of false-positive serological reaction between dengue and coronavirus disease 2019(COVID-19)and its implications for diagnosis.Evidence syntheses were conducted by systematically reviewing available literature using multiple databases,including Web of Science,PubMed,Google Scholar and medRxiv.Among a total of 16 presented cases from clinical settings,cross-reaction to COVID-19 serological tests was observed in two(12.5%)dengue-positive patients,while 14 patients(87.5%)confirmed positive for severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)showed a cross-reaction with dengue serological tests,leading to misdiagnosis and mismanagement by attending clinicians.Of 1789 SARS-CoV-2-positive sera,cross-reaction to dengue serological tests was observed in 180 sera(10%),which is higher than the cross-reaction observed for SARS-CoV-2 in archived pre-COVID-19 sera positive for a dengue infection(75 of 811,9.2%,P=0.674).Clinicians in tropical regions are therefore advised to interpret serological tests with caution and use a more pragmatic approach to triage these infections.
文摘In patients with respiratory allergy,cross-reactivity between aeroallergens and foods may induce food allergy,symptoms ranging from oral allergy syndrome to severe anaphylaxis.Clinical entities due to Ig E sensitization to cross-reactive aeroallergen and food allergen components are described for many sources of plant origin(pollen-food syndromes and associations,such as birch-apple,cypress-peach and celery-mugwortspice syndromes,and mugwort-peach,mugwortchamomile,mugwort-mustard,ragweed-melon-banana,goosefoot-melon associations),fungal origin(Alternariaspinach syndrome),and invertebrate,mammalian or avian origin(mite-shrimp,cat-pork,and bird-egg syndromes).Clinical cases of allergic reactions to ingestion of food products containing pollen grains of specific plants,in patients with respiratory allergy to Asteraceae pollen,especially mugwort and ragweed,are also mentioned,for honey,royal jelly and bee polen dietary supplements,along with allergic reactions to foods contaminated with mites or fungi in patients with respiratory allergy to these aeroallergens.Medical history and diagnosis approach may be guided by the knowledge about the diverse cross-reacting allergens involved,and by the understanding of these clinical entities which may vary significantly or may be overlapping.The association between primary Ig E sensitization with respiratory symptoms to inhaled allergens and food allergy due to cross-reactive allergen components is important to assess in allergy practice.The use of molecular-based diagnosis improves the understanding of clinically relevant Ig E sensitization to cross-reactive allergen components from aeroallergen sources and foods.
基金Supported by The National Natural Science Foundation of China, No 30500476The National High-Tech Science Foundation of China, No 2008AA02Z434+1 种基金National S and T Major Projects for Infectious Disease Control, No 2008ZX10002-013 and 2009ZX09103-621Beijing Natural Science Foundation, No 7082048
文摘AIM:To evaluate the presence and cross-reactive anti-bodies against hypervariable region 1(HVR1) in hepatitis C virus(HCV) infected patients and its relationship with the progression of the disease.METHODS:Sixteen representative HVR1 proteins selected from a unique set of 1600 natural sequences were used to semiquantitate the cross-reactivity of HVR1 antibodies in the sera of HCV patients.Fifty-five chronic HCV patients including 23 with asymptomatic mild hepatitis,18 with chronic hepatitis and 16 with liver cirrhosis patients were studied.RESULTS:The degree of the cross-reactivity of anti-HVR1 antibodies in 23 patients with mild asymptomatic hepatitis was 3.09 ± 2.68,which was signif icantly lower than in those with chronic hepatitis(5.44 ± 3.93,P < 0.05) and liver cirrhosis(7.44 ± 3.90,P < 0.01).No correlation was observed between the broadness of the cross-reactivity anti-HVR1 antibodies and patient's age,infection time,serum alanine aminotransferase activity,or serum HCV-RNA concentration.It was the breath of cross-reactivity rather than the presence of anti-HVR1 antibody in HCV sera that was associated with the progression of liver disease.CONCLUSION:The broadly cross-reactive HVR1 antibodies generated in natural HCV patients can not neutralize the virus,which results in persistent infection in patients with chronic hepatitis.
基金supported by the Natural Science Foundation of Guangdong Province,China(994162).
文摘An ester activation method was employed to couple enrofloxacin(ENFX) to the carrier proteins BSA and OVA. The conjugates ENFX-BSA and ENFX-OVA were identified with an UV spectrophotometer and amino acid automation analysis instrument, and resulted in conjugates with 48 ENFX molecules per carrier molecule(BSA). Splenocytes from mice immunized with ENFX-BSA were fused with SP2/0 myeloma cells and hybridomas secreting antibodies against enrofloxacin were selected and cloned. Two stable monoclonal antibodies, 2C5, 5D5 of the subclass IgG2a, were isolated. Using antibody 5D5, an indirect competitive inhibition enzyme-linked immunosorbent assay (Ci-ELISA) was developed for the quantitative detection of enrofloxacin and its metabolites. The IC50 of the standard curve was 21.67 ng mL-1 and the limit of detection for enrofloxacin was 0.13 ng mL-1. This method was sensitive and had a linear range from 0.13 to 10 000 ng mL-1 (r= -0.9782). Monoclonal antibody 5D5 exhibited high relative affinity to enrofloxacin, and the cross-reactivities with ciprofloxacin, marbofloxacin, sarafloxacin and danorfloxacin were 110.8, 27.40, 71.05 and 37.41%, respectively. Three non-fluoroquinolones of cefadroxil, chloramphenicol, sulfadimethoxine were tested and there was no cross-reaction between them.
基金supported by National Science and Technology Major Project of China[2018ZX10731301-002]。
文摘Objectives To evaluate the immunogenicity of Mycobacterium intracellulare proteins and determine the cross-reactive proteins between M.intracellulare and M.tuberculosis.Methods Protein extracts from M.intracellulare were used to immunize BALB/c mice.The antigens were evaluated using cellular and humoral immunoassays.The common genes between M.intracellular and M.tuberculosis were identified using genome-wide comparative analysis,and cross-reactive proteins were screened using immunoproteome microarrays.Results Immunization with M.intracellulare proteins induced significantly higher levels of the cytokines interferon-γ(IFN-γ),interleukin-2(IL-2),interleukin-12(IL-12),interleukin-6(IL-6)and immunoglobulins IgG,IgG1,IgM,and IgG2a in mouse serum.Bone marrow-derived macrophages isolated from mice immunized with M.intracellulare antigens displayed significantly lower bacillary loads than those isolated from mice immunized with adjuvants.Whole-genome sequence analysis revealed 396 common genes between M.intracellulare and M.tuberculosis.Microchip hybridization with M.tuberculosis proteins revealed the presence of 478 proteins in the serum of mice immunized with M.intracellulare protein extracts.Sixty common antigens were found using both microchip and genomic comparative analyses.Conclusion This is the advanced study to investigate the immunogenicity of M.intracellulare proteins and the cross-reactive proteins between M.intracellulare and M.tuberculosis.The results revealed the presence of a number of cross-reactive proteins between M.intracellulare and M.tuberculosis.Therefore,this study provides a new way of identifying immunogenic proteins for use in tuberculosis vaccines against both M.intracellulare and M.tuberculosis in future.
