Piglet performance and immunity is determined by the parity of both the birth dam and the rearing dam

2013 ◽  
Vol 53 (1) ◽  
pp. 46 ◽  
Author(s):  
Y. J. Miller ◽  
A. M. Collins ◽  
D. Emery ◽  
D. J. Begg ◽  
R. J. Smits ◽  
...  
Keyword(s):  
Innate Immunity ◽  
Adaptive Immunity ◽  
Whole Blood ◽  
Disease Susceptibility ◽  
Immune Cell ◽  
Saline Control ◽  
Enzyme Linked Immunosorbent Assay ◽  
Humoral Immune ◽  
Pathogen Challenge ◽  
Specific Igg

Gilt progeny have lower weaning weights and higher post-weaning medication and mortality rates, indicating greater disease susceptibility, than do sow progeny. The present study aimed to identify explanatory innate or adaptive immunity differences between gilt and sow progeny and potential pre- or post-natal influences. Sixty-four dams were vaccinated twice pre-farrowing with tetanus toxoid (TT). Serum (pre-vaccination) and colostrum and/or milk samples were collected to determine concentrations of TT-specific immunoglobulin G (IgG), by using an enzyme-linked immunosorbent assay (ELISA). Piglets were removed from their birth dam before suckling and fostered (not to their birth dam) to form 16 gilt and 16 sow litters, with five gilt-born and five sow-born piglets per litter. Piglets were vaccinated at weaning (4 weeks old) with either TT or saline (control). Sera and whole blood were collected from three gilt-born and three sow-born piglets per litter at 2, 4 and 7 weeks of age. Innate immunity was assessed indirectly on whole blood using an interferon gamma (IFN-γ) immune cell stimulation assay and a phagocytic assay. Piglets were weighed at birth, 4, 10, 17 and 22 weeks of age. There was no difference (P > 0.05) in the concentration of TT-specific IgG in colostrum and milk from gilts and older-parity sows, suggesting a similar ability to transfer IgG antibodies to a novel antigen. Birth dam parity did not affect piglets’ TT-specific IgG concentrations pre-weaning (P > 0.05) suggesting similar ability to absorb passively acquired IgG. Sow-reared piglets, however, had lower (P < 0.05) concentrations of TT-specific IgG than did gilt-reared piglets, possibly due to haemodilution in the faster-growing sow progeny. Gilt-born progeny had a reduced IgG response post-weaning to TT vaccination relative to sow-born progeny (P < 0.05), indicating adaptive immunity differences. Birth dam parity did not affect (P > 0.05) innate immunity (number/responsiveness of cells). Rearing dam parity influenced phagocytic activity pre- and post-weaning (gilt-reared > sow-reared; P < 0.05), possibly due to increased pathogen challenge. Birthweight was affected by birth dam parity (sow-born > gilt-born; P < 0.05) while rearing dam parity determined weaning weight (sow-reared > gilt-reared; P < 0.05), with no difference evident at 22 weeks. The results of the present study suggest that gilt-born progeny may be more susceptible to disease post-weaning than sow-born progeny due to their lower birthweight and reduced humoral immune responsiveness. The rearing dam may also affect disease susceptibility in progeny due to slower pre-weaning growth, lower weaning weights and increased pathogen challenge, both pre- and post-weaning.

scholarly journals Immunogenicity of an Autogenous Streptococcus suis Bacterin in Preparturient Sows and Their Piglets in Relation to Protection after Weaning

2010 ◽  
Vol 17 (10) ◽  
pp. 1589-1597 ◽  
Author(s):  
Christoph Georg Baums ◽  
Christian Brüggemann ◽  
Christoph Kock ◽  
Andreas Beineke ◽  
Karl-Heinz Waldmann ◽  
...  
Keyword(s):  
Streptococcus Suis ◽  
Inhibitory Effect ◽  
Active Immunization ◽  
Enzyme Linked Immunosorbent Assay ◽  
Maternal Immunity ◽  
Humoral Immune ◽  
Specific Pathogen ◽  
Suckling Piglets ◽  
Specific Igg ◽  
Weaning Piglets

ABSTRACT Streptococcus suis is an important porcine pathogen causing meningitis and other invasive diseases in piglets of different ages. Application of S. suis serotype 2 bacterins to specific-pathogen-free (SPF) weaning piglets has been demonstrated to protect against the homologous serotype. However, autogenous S. suis bacterins are also applied to sows and suckling piglets in the field. Therefore, comparative evaluation of different bacterin immunization regimes, including sow vaccination, was performed in this study. The main objectives were to determine the immunogenicity of an S. suis bacterin in sows prepartum and its influence on active immunization of piglets. Experimental infection of 6- and 8-week-old weaning piglets was performed to elucidate protective efficacies. Humoral immune responses were investigated by an enzyme-linked immunosorbent assay (ELISA) measuring muramidase-released protein (MRP)-specific IgG titers and by opsonophagocytosis assays. Bacterin application elicited high MRP-specific IgG titers in the serum and colostrum of sows, as well as opsonizing antibodies. Piglets from vaccinated sows had significantly higher MRP-specific titers than respective piglets from nonvaccinated sows until 6 weeks postpartum. Vaccination of suckling piglets did not result in high MRP-specific titers nor in induction of opsonizing antibodies. Furthermore, neither vaccination of suckling nor of weaning piglets from immunized sows was associated with a prominent active immune response and protection at 8 weeks postpartum. However, protection was observed in respective 6-week-old weaning piglets, most likely because of protective maternal immunity. In conclusion, this study provides the first results suggesting protective passive maternal immunity for S. suis serotype 2 after bacterin vaccination of sows and a strong inhibitory effect on active immunization of suckling and weaning piglets, leading to highly susceptible growers.

scholarly journals Humoral and Cellular Immunogenicity of Zoster Vaccine within One Year after Herpes Zoster

2017 ◽  
Vol 4 (suppl_1) ◽  
pp. S414-S414
Author(s):  
Eunyoung Lee ◽  
June Young Chun ◽  
Kyoung-Ho Song ◽  
Pyeong Gyun Choe ◽  
Ji Whan Bang ◽  
...  
Keyword(s):  
Herpes Zoster ◽  
Geometric Mean ◽  
Enzyme Linked Immunosorbent Assay ◽  
Prior History ◽  
Zoster Vaccine ◽  
Varicella Zoster ◽  
Humoral Immune ◽  
History Of ◽  
Specific Igg ◽  
One Year

Abstract Background Herpes zoster vaccination is recommended to patients with a prior history of herpes zoster to prevent reactivation. However, the appropriate timing of vaccination is controversial. We compared immunogenicity of vaccine according to timing of vaccination after zoster illness. Methods In this prospective observational study, subjects were stratified into two groups by the vaccination timing since their zoster illness: 6–12 months (within-1 year group) vs. 1–5 years (after-1 year group). Blood samples were collected before and 6 weeks after vaccination of zoster vaccine live. Varicella-zoster virus (VZV)-specific IgG concentrations were measured by enzyme-linked immunosorbent assay. Interferon-gamma enzyme-linked immunosorbent spot (ELISPOT) assays were performed to assess VZV specific T-cell responses. Results A total of 59 patients (18 in the within-1 year group and 41 in the after-1 year group) were enrolled. Ages were not significantly different between groups. The baseline geometric mean titer (GMT) of VZV IgG was higher in the within-1 year group than in the after-1 year group (245.8 IU/mL vs. 124.9 IU/mL; P = 0.040). The geometric mean fold-rise (GMFR) of VZV IgG was lower in the within-1 year group than in the after-1 year group (1.42 vs. 2.46; P = 0.002). The GMT of spot forming cell (SFC) counts by ELISPOT at baseline and 6 weeks after vaccination were not significantly different between groups. The GMFRs of SFCs were also comparable. Conclusion Zoster vaccination within 1 year after zoster illness may have disadvantage in the aspect of humoral immune response (ClinicalTrials.gov number, NCT02704572). Disclosures All authors: No reported disclosures.

scholarly journals Bridging autoinflammatory and autoimmune diseases

2021 ◽  
Vol 33 (1) ◽  
Author(s):  
Emad M. El-Shebiny ◽  
Enas S. Zahran ◽  
Sabry A. Shoeib ◽  
Eman S. Habib
Keyword(s):  
Innate Immunity ◽  
T Lymphocytes ◽  
Autoimmune Diseases ◽  
Adaptive Immunity ◽  
High Titre ◽  
The Other ◽  
Clinical Spectrum ◽  
Autoinflammatory Diseases ◽  
Main Body ◽  
B And T Lymphocytes

Abstract Background Autoimmunity is used to cause by impairment of adaptive immunity alone, whereas autoinflammatory was originally defined as a consequence of unregulated innate immunity. So, the pathogenetic mechanisms of autoimmune diseases were well-thought-out to be mediated by B and T lymphocytes. Whereas, autoinflammatory diseases were defined as unprovoked times of inflammation with the absence of a high titre of autoantibodies. Main body of the abstract Autoimmune and autoinflammatory diseases were split into two groups, but considering the similarities, it can be considered as only one group of diseases with a large immune pathological and clinical spectrum which involves at one end pure autoimmune diseases and the other pure autoinflammatory diseases. Conclusions We can safely conclude that there is bridging between autoinflammatory and autoimmune diseases.

scholarly journals OP0286 PROTEOMICS AND RNA SEQUENCING APPROACHES HIGHLIGHT THE ROLE OF ENDOTHELIAL CELL DYSREGULATION IN IL-1 AND IFN MEDIATED AUTOINFLAMMATORY DISEASES, NOMID AND CANDLE

2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 178-179
Author(s):  
S. Alehashemi ◽  
M. Garg ◽  
B. Sellers ◽  
A. De Jesus ◽  
A. Biancotto ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Whole Blood ◽  
Cell Activation ◽  
Immune Cell ◽  
Differentially Expressed ◽  
Neutrophilic Dermatosis ◽  
Healthy Controls ◽  
Differentially Expressed Proteins ◽  
Rna Seq ◽  
Before And After

Background:Systemic Autoinflammatory diseases present with sterile inflammation. NOMID (Neonatal-Onset Multisystem Inflammatory Disease) is caused by gain-of-function mutations inNLRP3and excess IL-1 production, presents with fever, neutrophilic dermatosis, aseptic meningitis, hearing loss and eye inflammation; CANDLE (Chronic Atypical Neutrophilic Dermatosis, Lipodystrophy and Elevated Temperature) is caused by loss-of-function mutations in proteasome genes that lead to type-1 interferon signaling, characterized by fever, panniculitis, lipodystrophy, cytopenia, systemic and pulmonary hypertension and basal ganglia calcification. IL-1 blockers are approved for NOMID and JAK-inhibitors show efficacy in CANDLE treatment.Objectives:We used proteomic analysis to compare differentially expressed proteins in active NOMID and CANDLE compared to healthy controls before and after treatment, and whole blood bulk RNA seq to identify the immune cell signatures.Methods:Serum samples from active NOMID (n=12) and CANDLE (n=7) before and after treatment (table 1) and age matched healthy controls (HC) (n=7) were profiled using the SomaLogic platform (n=1125 proteins). Differentially expressed proteins in NOMID and CANDLE were ranked after non-parametric tests for unpaired (NOMIDp<0.05, CANDLE,p<0.1) and paired (p<0.05) analysis and assessed by enriched Gene Ontology pathways and network visualization. Whole blood RNA seq was performed (NOMID=7, CANDLE=7, Controls =5) and RPKM values were used to assess immune cells signatures.Table 1.Patient’s characteristicsNOMIDN=12, Male =6CANDLEN=7, Male =6AgeMedian (range)12 (2, 28)16 (3, 20)Ethnicity%White (Hispanic)80 (20)100 (30)GeneticsNLRP3mutation(2 Somatic, 10 Germline)mutations in proteasome component genes(1 digenic, 6 Homozygous/compound Heterozygous)Before treatmentAfter treatmentBefore treatmentAfter treatmentCRPMedian (range) mg/L52 (16-110)5 (0-23)5 (0-101)1 (0-4)IFN scoremedian (range)0NA328 (211-1135)3 (0-548)Results:Compared to control, 205 proteins (127 upregulated, 78 downregulated) were significantly different at baseline in NOMID, compared to 163 proteins (101 upregulated, and 62 downregulated) in CANDLE. 134 dysregulated proteins (85 upregulated, 49 downregulated) overlapped in NOMID and CANDLE (Figure 1). Pathway analysis identified neutrophil and monocyte chemotaxis signature in both NOMID and CANDLE. NOMID patients had neutrophilia and active neutrophils. CANDLE patients exhibited active neutrophils in whole blood RNA. Endothelial cell activation was the most prominent non-hematopoietic signature and suggest distinct endothelial cell dysregulation in NOMID and CANDLE. In NOMID, the signature included neutrophil transmigration (SELE) endothelial cell motility in response to angiogenesis (HGF, VEGF), while in CANDLE the endothelial signatures included extracellular matrix protein deposition (COL8A) suggesting increased vascular stiffness. CANDLE patients had higher expression of Renin, 4 out of 7 had hypertension, NOMID patients did not have hypertension. Treatment with anakinra and baricitinib normalized 143 and 142 of dysregulated proteins in NOMID and CANDLE respectively.Conclusion:Differentially expressed proteins in NOMID and CANDLE are consistent with innate immune cell activation. Distinct endothelial cell signatures in NOMID and CANDLE may provide mechanistic insight into differences in vascular phenotypes. Treatment with anakinra and Baricitinib in NOMID and CANDLE leaves 30% and 13% of the dysregulated proteins unchanged.Acknowledgments:This work was supported by Intramural Research atNational Institute of Allergy Immunology and Infectious Diseases of National Institutes of Health, Bethesda, Maryland, the Center of Human Immunology and was approved by the IRB.Disclosure of Interests:None declared

scholarly journals Injury-Induced Innate Immune Response During Segment Regeneration of the Earthworm, Eisenia andrei

2021 ◽  
Vol 22 (5) ◽  
pp. 2363
Author(s):  
Kornélia Bodó ◽  
Zoltán Kellermayer ◽  
Zoltán László ◽  
Ákos Boros ◽  
Bohdana Kokhanyuk ◽  
...  
Keyword(s):  
Immune Response ◽  
Innate Immunity ◽  
Immune Cell ◽  
Scavenger Receptor ◽  
Cell Renewal ◽  
Body Parts ◽  
Eisenia Andrei ◽  
Blastema Formation ◽  
Close Proximity ◽  
Renewal Period

Regeneration of body parts and their interaction with the immune response is a poorly understood aspect of earthworm biology. Consequently, we aimed to study the mechanisms of innate immunity during regeneration in Eisenia andrei earthworms. In the course of anterior and posterior regeneration, we documented the kinetical aspects of segment restoration by histochemistry. Cell proliferation peaked at two weeks and remitted by four weeks in regenerating earthworms. Apoptotic cells were present throughout the cell renewal period. Distinct immune cell (e.g., coelomocyte) subsets were accumulated in the newly-formed blastema in the close proximity of the apoptotic area. Regenerating earthworms have decreased pattern recognition receptors (PRRs) (e.g., TLR, except for scavenger receptor) and antimicrobial peptides (AMPs) (e.g., lysenin) mRNA patterns compared to intact earthworms. In contrast, at the protein level, mirroring regulation of lysenins became evident. Experimental coelomocyte depletion caused significantly impaired cell divisions and blastema formation during anterior and posterior regeneration. These obtained novel data allow us to gain insight into the intricate interactions of regeneration and invertebrate innate immunity.

scholarly journals SARS-CoV-2 specific antibody and neutralization assays reveal the wide range of the humoral immune response to virus

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Mikail Dogan ◽  
Lina Kozhaya ◽  
Lindsey Placek ◽  
Courtney Gunter ◽  
Mesut Yigit ◽  
...  
Keyword(s):  
Specific Antibody ◽  
Neutralizing Antibodies ◽  
Vaccine Development ◽  
High Specificity ◽  
Spike Protein ◽  
Humoral Immune ◽  
Specificity And Sensitivity ◽  
Wide Range ◽  
Specific Igg ◽  
Negative Controls

AbstractDevelopment of antibody protection during SARS-CoV-2 infection is a pressing question for public health and for vaccine development. We developed highly sensitive SARS-CoV-2-specific antibody and neutralization assays. SARS-CoV-2 Spike protein or Nucleocapsid protein specific IgG antibodies at titers more than 1:100,000 were detectable in all PCR+ subjects (n = 115) and were absent in the negative controls. Other isotype antibodies (IgA, IgG1-4) were also detected. SARS-CoV-2 neutralization was determined in COVID-19 and convalescent plasma at up to 10,000-fold dilution, using Spike protein pseudotyped lentiviruses, which were also blocked by neutralizing antibodies (NAbs). Hospitalized patients had up to 3000-fold higher antibody and neutralization titers compared to outpatients or convalescent plasma donors. Interestingly, some COVID-19 patients also possessed NAbs against SARS-CoV Spike protein pseudovirus. Together these results demonstrate the high specificity and sensitivity of our assays, which may impact understanding the quality or duration of the antibody response during COVID-19 and in determining the effectiveness of potential vaccines.

scholarly journals Immunological Aspects of SARS-CoV-2 Infection and the Putative Beneficial Role of Vitamin-D

2021 ◽  
Vol 22 (10) ◽  
pp. 5251
Author(s):  
Ming-Yieh Peng ◽  
Wen-Chih Liu ◽  
Jing-Quan Zheng ◽  
Chien-Lin Lu ◽  
Yi-Chou Hou ◽  
...  
Keyword(s):  
Vitamin D ◽  
Innate Immunity ◽  
T Cells ◽  
Adaptive Immunity ◽  
Cd4 T Cells ◽  
Vitamin D Supplementation ◽  
The Body ◽  
Alveolar Type ◽  
Tissue Destruction ◽  
Health Crisis

Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is still an ongoing global health crisis. Immediately after the inhalation of SARS-CoV-2 viral particles, alveolar type II epithelial cells harbor and initiate local innate immunity. These particles can infect circulating macrophages, which then present the coronavirus antigens to T cells. Subsequently, the activation and differentiation of various types of T cells, as well as uncontrollable cytokine release (also known as cytokine storms), result in tissue destruction and amplification of the immune response. Vitamin D enhances the innate immunity required for combating COVID-19 by activating toll-like receptor 2. It also enhances antimicrobial peptide synthesis, such as through the promotion of the expression and secretion of cathelicidin and β-defensin; promotes autophagy through autophagosome formation; and increases the synthesis of lysosomal degradation enzymes within macrophages. Regarding adaptive immunity, vitamin D enhances CD4+ T cells, suppresses T helper 17 cells, and promotes the production of virus-specific antibodies by activating T cell-dependent B cells. Moreover, vitamin D attenuates the release of pro-inflammatory cytokines by CD4+ T cells through nuclear factor κB signaling, thereby inhibiting the development of a cytokine storm. SARS-CoV-2 enters cells after its spike proteins are bound to angiotensin-converting enzyme 2 (ACE2) receptors. Vitamin D increases the bioavailability and expression of ACE2, which may be responsible for trapping and inactivating the virus. Activation of the renin–angiotensin–aldosterone system (RAS) is responsible for tissue destruction, inflammation, and organ failure related to SARS-CoV-2. Vitamin D inhibits renin expression and serves as a negative RAS regulator. In conclusion, vitamin D defends the body against SARS-CoV-2 through a novel complex mechanism that operates through interactions between the activation of both innate and adaptive immunity, ACE2 expression, and inhibition of the RAS system. Multiple observation studies have shown that serum concentrations of 25 hydroxyvitamin D are inversely correlated with the incidence or severity of COVID-19. The evidence gathered thus far, generally meets Hill’s causality criteria in a biological system, although experimental verification is not sufficient. We speculated that adequate vitamin D supplementation may be essential for mitigating the progression and severity of COVID-19. Future studies are warranted to determine the dosage and effectiveness of vitamin D supplementation among different populations of individuals with COVID-19.

scholarly journals Duck RIG-I CARD Domain Induces the Chicken IFN-βby Activating NF-κB

2015 ◽  
Vol 2015 ◽  
pp. 1-6 ◽  
Author(s):  
Yang Chen ◽  
Zhengyang Huang ◽  
Bin Wang ◽  
Qinming Yu ◽  
Ran Liu ◽  
...  
Keyword(s):  
Innate Immunity ◽  
Eukaryotic Expression ◽  
Poly I:C ◽  
Expression Vectors ◽  
Luciferase Reporter ◽  
Enzyme Linked Immunosorbent Assay ◽  
Regulatory Domains ◽  
Polycytidylic Acid ◽  
Card Domain ◽  
Inducible Gene

Retinoic acid-inducible gene I- (RIG-I-) like receptors (RLRs) have recently been identified as cytoplasmic sensors for viral RNA. RIG-I, a member of RLRs family, plays an important role in innate immunity. Although previous investigations have proved that RIG-I is absent in chickens, it remains largely unknown whether the chicken can respond to RIG-I ligand. In this study, the eukaryotic expression vectors encoding duRIG-I full length (duck RIG-I, containing all domains), duRIG-I N-terminal (containing the two caspase activation and recruitment domain, CARDs), and duRIG-I C-terminal (containing helicase and regulatory domains) labeled with 6*His tags were constructed successfully and detected by western blotting. Luciferase reporter assay and enzyme-linked immunosorbent assay (ELISA) detected the duRIG-I significantly activated NF-κB and induced the expression of IFN-βwhen polyinosinic-polycytidylic acid (poly[I:C], synthetic double-stranded RNA) challenges chicken embryonic fibroblasts cells (DF1 cells), while the duRIG-I was inactive in the absence of poly[I:C]. Further analysis revealed that the CARDs (duRIG-I-N) induced IFN-βproduction regardless of the presence of poly[I:C], while the CARD-lacking duRIG-I (duRIG-I-C) was not capable of activating downstream signals. These results indicate that duRIG-I CARD domain plays an important role in the induction of IFN-βand provide a basis for further studying the function of RIG-I in avian innate immunity.

scholarly journals Characterization of the Humoral Immune Response to Chlamydia Outer Membrane Protein 2 in Chlamydial Infection

2003 ◽  
Vol 10 (1) ◽  
pp. 103-107 ◽  
Author(s):  
I. Portig ◽  
J. C. Goodall ◽  
R. L. Bailey ◽  
J. S. H. Gaston
Keyword(s):  
Membrane Protein ◽  
Outer Membrane ◽  
Outer Membrane Protein ◽  
Chlamydial Infection ◽  
Chlamydia Pneumoniae ◽  
Specific Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Recent Infection ◽  
Humoral Immune ◽  
Immunodominant Antigen

ABSTRACT Detection of antibodies to an outer membrane protein 2 (OMP2) by enzyme-linked immunosorbent assay (ELISA) by using either the Chlamydia trachomatis- or the Chlamydia pneumoniae-specific protein was investigated. OMP2 is an immunodominant antigen giving rise to antibody responses in humans infected with different C. trachomatis serovars (A to C and D to K) or with C. pneumoniae, which could be detected by OMP2 ELISA. OMP2 ELISA is not species specific, but antibody titers were usually higher on the homologous protein. The sensitivity of this assay was high but varied according to the “gold standard” applied. Levels of antibody to C. pneumoniae OMP2 as detected by ELISA seem to return to background or near-background values within a shorter period of time compared to antibodies to C. pneumoniae detected by microimmunofluorescence (MIF), making it more likely that positive results in ELISA reflect recent infection. Thus, OMP2 ELISA has distinct advantages over MIF and commercially available ELISAs and might be a useful tool for the serodiagnosis of chlamydial infection.

scholarly journals Purified Hexameric Epstein-Barr Virus-Encoded BARF1 Protein for Measuring Anti-BARF1 Antibody Responses in Nasopharyngeal Carcinoma Patients

2010 ◽  
Vol 18 (2) ◽  
pp. 298-304 ◽  
Author(s):  
E. K. Hoebe ◽  
S. H. Hutajulu ◽  
J. van Beek ◽  
S. J. Stevens ◽  
D. K. Paramita ◽  
...  
Keyword(s):  
Nasopharyngeal Carcinoma ◽  
Epstein Barr Virus ◽  
Antibody Responses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Humoral Immune ◽  
Barr Virus ◽  
Humoral Immune Responses ◽  
Human Sera ◽  
Epstein Barr ◽  
Humoral Responses

ABSTRACTWHO type III nasopharyngeal carcinoma (NPC) is highly prevalent in Indonesia and 100% associated with Epstein-Barr virus (EBV). NPC tumor cells express viral proteins, including BARF1, which is secreted and is considered to have oncogenic and immune-modulating properties. Recently, we found conserved mutations in the BARF1 gene in NPC isolates. This study describes the expression and purification of NPC-derived BARF1 and analyzes humoral immune responses against prototype BARF1 (B95-8) and purified native hexameric BARF1 in sera of Indonesian NPC patients (n= 155) compared to healthy EBV-positive (n= 56) and EBV-negative (n= 16) individuals. BARF1 (B95-8) expressed inEscherichia coliand baculovirus, as well as BARF1-derived peptides, did not react with IgG or IgA antibodies in NPC. Purified native hexameric BARF1 protein isolated from culture medium was used in enzyme-linked immunosorbent assay (ELISA) and revealed relatively weak IgG and IgA responses in human sera, although it had strong antibody responses to other EBV proteins. Higher IgG reactivity was found in NPC patients (P= 0.015) than in regional Indonesian controls or EBV-negative individuals (P< 0.001). IgA responses to native BARF1 were marginal. NPC sera with the highest IgG responses to hexameric BARF1 in ELISA showed detectable reactivity with denatured BARF1 by immunoblotting. In conclusion, BARF1 has low immunogenicity for humoral responses and requires native conformation for antibody binding. The presence of antibodies against native BARF1 in the blood of NPC patients provides evidence that the protein is expressed and secreted as a hexameric protein in NPC patients.

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