BACKGROUND: The placenta serves as a vital interface between mother and fetus, facilitating nutrient exchange and immune regulation. Although generally effective in preventing pathogen transmission, some viruses such as...BACKGROUND: The placenta serves as a vital interface between mother and fetus, facilitating nutrient exchange and immune regulation. Although generally effective in preventing pathogen transmission, some viruses such as CMV and Zika can cross this barrier. The mechanism of vertical transmission of SARS-CoV-2 remains unclear. OBJECTIVE: To investigate the expression and interaction of caveolin-1 (Cav-1) and endothelial nitric oxide synthase (eNOS) in placental tissues from SARS-CoV-2-positive pregnancies and their potential role in limiting vertical transmission. MATERIALS AND METHODS: Placental samples were obtained from RT-PCR-confirmed COVID-19-positive and healthy pregnancies. Histological, immunohistochemical, and immunofluorescence techniques were used to evaluate tissue morphology and Cav-1/eNOS expression. RESULTS: Control placentas exhibited intact villous structures, normal erythrocyte morphology, and strong Cav-1 and eNOS expression in endothelial cells. SARS-CoV-2-positive placentas showed villous damage, swollen erythrocytes, decreased collagen, and reduced endothelial expression of Cav-1 and eNOS. Cav-1 was absent in syncytiotrophoblasts in all samples. CONCLUSION: Reduced expression of Cav-1 and eNOS in endothelial cells, combined with the absence of Cav-1 in syncytiotrophoblasts, may contribute to the placental resistance against SARS-CoV-2. These findings support the hypothesis that molecular features of the placenta, particularly involving Cav-1/eNOS pathways, play a protective role in preventing vertical transmission.
Piazza A, Rombolà G, Maggiore U
… +10 more, Ciappi D, Cozzi E, de Stefano MC, Crocchiolo R, Ricci A, Cardillo M, Miotti V, Feltrin G, Papola F, AIBT Working Group
Reliable and reproducible testing of Donor Specific HLA-Antibodies (DSA) is essential for clinical decisions in organ transplantation. In this prospective multicentre study from the Italian Transplant Network, we assesse...Reliable and reproducible testing of Donor Specific HLA-Antibodies (DSA) is essential for clinical decisions in organ transplantation. In this prospective multicentre study from the Italian Transplant Network, we assessed reproducibility and possible bias of Mean Fluorescence Intensity (MFI) values of Luminex-Single Antigen Bead (SAB) assays across 20 laboratories using the 2 commercially available platforms (Vendor 1 and Vendor 2). Both Vendors showed inter-laboratory median MFI coefficient of variation below 20 %. We evaluated the possible MFI bias between Vendors. In the clinically significant 1000-10000 MFI range, a strong MFI bias was observed for class I SAB, with Vendor 1 MFI being approximately one third of Vendor 2 (Vendor 1 MFI = 0.36 Vendor 2 MFI). For class II, the MFI bias was lower, approximately three quarters of Vendor 2 (Vendor 1 MFI = 0.74 Vendor 2 MFI), with several discrepant results for DQ alleles. Ultimately, we defined the best fitting cutoff values for positive and permissive DSA (1000 and 3000, respectively) in use in the current Italian National Kidney Allocation System. These were 500/1000 and 500/2000 for class I and class II for Vendor 1 compared to 1000/3000 for both classes in Vendor 2. Overall, we developed a tool to compare MFI values between different platforms for consistent DSA assignment across laboratories and enable the most appropriate clinical decisions in organ transplantation.
BACKGROUND: Tetanus prevention is effectively managed in infants and young children, yet there remains a need for enhanced measures to improve tetanus prevention among adults. METHODS: This phase IV, single-center, rando...BACKGROUND: Tetanus prevention is effectively managed in infants and young children, yet there remains a need for enhanced measures to improve tetanus prevention among adults. METHODS: This phase IV, single-center, randomized, double-blind, positive-controlled trial evaluated an adsorbed tetanus vaccine in individuals aged ≥ 16 years, stratified into 16-18 years, 19-49 years, and ≥ 50 years cohorts. RESULTS: Pre-vaccination seroprotection rates (SPR, ≥0.1 IU/mL) demonstrated age-dependent declines (44.00 %→18.47 %→5.38 %), paralleled by decreasing geometric mean concentrations (GMC: 0.079 IU/mL → 0.052 IU/mL → 0.033 IU/mL). A single booster dose achieved universal seroprotection (100 % SPR) and near-complete seroconversion (SCR ≥ 99.48 %) across all groups, elevating GMCs to 6.831 IU/mL (16-18 years), 3.400 IU/mL (19-49 years), and 1.430 IU/mL (≥50 years), while two doses in the ≥ 50 years cohort further increased GMC to 2.938 IU/mL with 100 % SCR. The vaccine exhibited a favorable safety profile, with mild vaccination-site pain (7.2 %) and pruritus (4.2 %) as predominant reactions, no vaccine-related serious adverse events, and self-limiting systemic effects (pyrexia: 1.6 %; myalgia: 1.2 %). CONCLUSION: These results confirm robust immunogenicity and safety in adults, supporting tailored booster strategies for aging populations with waning immunity.
Autoimmune diseases occur when the immune system mistakenly attacks healthy tissues, leading to chronic inflammation and progressive organ damage. These conditions affect millions worldwide and often result in long-term...Autoimmune diseases occur when the immune system mistakenly attacks healthy tissues, leading to chronic inflammation and progressive organ damage. These conditions affect millions worldwide and often result in long-term disability. The underlying reasons for increased vulnerability remain uncertain, particularly regarding the combined influence of genetic variations and lifestyle factors on disease risk. The purpose of this study is to identify key genetic variants associated with rheumatoid arthritis, systemic lupus erythematosus, multiple sclerosis, and type 1 diabetes, and to investigate how these variants interact with environmental exposures to influence risk and age of onset. Publicly available genetic and molecular data were analyzed, including genome-wide association studies (NHGRI-EBI GWAS Catalog, ImmunoBase, and dbGaP), DNA sequencing datasets (1000 Genomes Project, Genomic Data Commons, and SRA), gene expression profiles (GEO and ArrayExpress), and epigenetic marks (ENCODE and RegulomeDB). This study identifies several significant genetic variants associated with autoimmune diseases, including PTPN22, HLA-DRB1, HLA-DQA1, IRF5, IL10, and other immune-regulatory genes. Notably, the PTPN22 rs2476601 variant increased the risk by approximately 1.8 times, while the HLA-DRB1 rs3135388 variant increased the risk by around 2.1 times. Additional variants, such as HLA-DQ, IRF, and IL10, also play significant roles in immune dysregulation and susceptibility to autoimmune diseases, with specific allele associations yet to be fully explored. Pathway analysis revealed that these genes are primarily involved in antigen presentation, cytokine signaling, and T-cell activation-key processes underlying the pathogenesis of autoimmune diseases. Smoking and vitamin D deficiency further elevated risk in genetically susceptible individuals and were linked with earlier disease onset. The findings highlight genetic markers such as IL2RA and TIM-3 as potential tools for early screening and therapeutic targeting. Integrating genetic testing with lifestyle assessment can support personalized prevention, guide drug development, and improve clinical practice. Validation in diverse populations and long-term studies is recommended to confirm clinical usefulness.
Psoriasis (PsO) is a chronic, immune-mediated inflammatory disease with systemic manifestations. Its etiology involves an interaction between genetic, immunologic, and environmental factors. The NLRP3 inflammasome plays...Psoriasis (PsO) is a chronic, immune-mediated inflammatory disease with systemic manifestations. Its etiology involves an interaction between genetic, immunologic, and environmental factors. The NLRP3 inflammasome plays a key role in innate immune responses and has been implicated in PsO pathogenesis. However, data regarding NLRP3 gene polymorphisms and their relationship with PsO, particularly in the Brazilian population, remain limited. The aim was to investigate the association between the NLRP3 rs10157379 and rs10754558 gene variants with susceptibility and severity of PsO, as well as the risk of developing psoriatic arthritis (PsA). A case-control study was conducted with 154 individuals with PsO and 154 healthy controls. Clinical and demographic data were collected, and genotyping of the NLRP3 rs10157379 T > C and rs10754558 G > C variants was performed using real-time polymerase chain reaction. No significant association was observed between either variant and PsO susceptibility or severity. However, the rs10754558 CG genotype was associated with a reduced likelihood of PsA [odds ratio (OR): 0.32; 95 % confidence interval (CI): 0.10-0.95, p = 0.041], while the GG genotype conferred a higher likelihood (OR: 3.26; 95 % CI:1.03-10.32, p = 0.044). No associations were identified for the rs10157379 variant. In conclusion, the NLRP3 rs10157379 and rs10754558 variants were not associated with PsO susceptibility or severity. However, the rs10754558 variant may influence the chance of joint involvement in individuals with PsO. These findings underscore the potential role of NLRP3 variants in PsA development and highlight the importance of genetic background in PsO pathophysiology.
Reduction in pre-transplant HLA donor specific antibodies (DSA) is a known phenomenon after liver or simultaneous liver kidney (SLK) transplants. However, kidney after liver transplant may be associated with increased im...Reduction in pre-transplant HLA donor specific antibodies (DSA) is a known phenomenon after liver or simultaneous liver kidney (SLK) transplants. However, kidney after liver transplant may be associated with increased immunologic risk, in highly sensitized patients. There is no consensus in avoidance of antibodies in patients with strong historical anti-HLA antibodies who require a second organ after liver transplant. We present a case series of 4 highly sensitized liver recipients who needed a safety net kidney transplant (snKT). To determine which antigens should be avoided, detailed eplet analysis of pre and post liver transplant antibody showed that the liver had successfully cleared eplet-associated DSA; however, third party eplet antibodies persisted. The liver's role in the clearance of DSA versus third party antibody is inconclusive due to a lack of granular eplet analysis data in the literature. Herein, eplet analysis allowed for more accurate determination of higher risk antibodies for snKT.
Eplet mismatch (MM) quantification is a valuable tool for refining histocompatibility in kidney transplantation, especially at HLA-DR/DQ loci. However, zero MM at these loci does not ensure complete immunological safety....Eplet mismatch (MM) quantification is a valuable tool for refining histocompatibility in kidney transplantation, especially at HLA-DR/DQ loci. However, zero MM at these loci does not ensure complete immunological safety. We report a 17-year-old male who received a living donor kidney transplant via the kidney paired donation (KPD) program. The donor had 0 eplet MMs at HLA-DR/DQ, with negative crossmatch and no pre-transplant donor-specific antibodies (DSAs). Despite this, the patient developed early mixed rejection-T cell-mediated (TCMR) and antibody-mediated (ABMR), and further analysis revealed DSAs targeting an HLA-C 80 K/77 N eplet. Combined treatment for TCMR and ABMR led to histological and functional improvement. This case illustrates the limitation of relying solely on HLA-DR/DQ eplet matching. Despite achieving 0 MM at these loci, the patient developed significant alloimmune injury. Molecular MM scores are useful predictive biomarkers, but they do not fully capture the complexity of immune responses in transplantation.
This study investigated the prevalence of BKV subtypes in kidney transplant recipients and evaluates their association with viral load and BKVAN risk. A total of 32 kidney transplant recipients with confirmed BKV infecti...This study investigated the prevalence of BKV subtypes in kidney transplant recipients and evaluates their association with viral load and BKVAN risk. A total of 32 kidney transplant recipients with confirmed BKV infection were included in the study. BKV DNA was extracted from urine and plasma samples and genotyped using phylogenetic analysis and a novel genotyping algorithm. Viral loads were compared between subtypes, and the occurrence of BKVAN was analyzed in relation to genotype distribution All samples belonged to genotype I, with 78.5 % classified as Ib-2 and 21.4 % as Ib-1. No genotype II, III, or IV strains were detected. Although the Ib-1 subtype exhibited higher mean viral loads in both urine and plasma, the differences were not statistically significant (p = 0.055 and p = 0.07, respectively). Two patients developed BKVAN, both infected with the Ib-2 subtype. The genotyping algorithm demonstrated a 98 % concordance rate with phylogenetic analysis, confirming its reliability. The predominance of BKV genotype Ib-2 among kidney transplant recipients aligns with global trends. While no significant association between genotype and viral load was found, all BKVAN cases were linked to the Ib-2 subtype, suggesting a potential role in disease progression.
A 31-year-old male undergoing evaluation for an allogeneic stem cell transplant exhibited a complete mismatch at the MICA locus compared to his biological mother. The recipient was homozygous MICA*008:04, while the mothe...A 31-year-old male undergoing evaluation for an allogeneic stem cell transplant exhibited a complete mismatch at the MICA locus compared to his biological mother. The recipient was homozygous MICA*008:04, while the mother was homozygous MICA*002:01. Sample mix-up was ruled out and we sought an alternative explanation. Upon investigation of MICA and HLA-B allele linkage, multiple studies reported an association of a MICA deletion (MICAdel) with HLA-B*48. Interestingly, the patient and his mother both typed as HLA-B*48:02. Furthermore, the patient and his mother each typed as MICB*09:01N. Copy number analysis confirmed a single copy of MICA in both samples, thus providing supporting evidence of a MICAdel in both subjects, and consistent with previously published data on the association of a MICAdel with HLA-B*48. What initially appeared to be a maternal exclusion based on a MICA mismatch was instead attributable to a hemizygous loss of MICA. This case highlights two important considerations: 1) evaluating other loci, not generally considered in most clinical matching algorithms can uncover interesting genetic anomalies usually described only in research settings, 2) The appreciation of such findings can lead to studies that explore the clinical impact of such an evolutionary change.
Umbilical cord blood (UCB) is a valuable source of hematopoietic stem cells for treating life-threatening haematological disorders. India's vast population, known for its cultural, linguistic, and genetic diversity, requ...Umbilical cord blood (UCB) is a valuable source of hematopoietic stem cells for treating life-threatening haematological disorders. India's vast population, known for its cultural, linguistic, and genetic diversity, requires a significant need for HLA-matched donors. We analyzed HLA-A, -B, -C, -DRB1, and -DQB1 allele and haplotype diversity in 30,027 UCB units. Samples were categorized into six broad regional populations and further sub-clustered into nine groups based on the predominant language spoken. The most frequent haplotypes identified were A*33:03 ∼ B*44:03 ∼ C*07:06 ∼ DRB1*07:01 ∼ DQB1*02:02 (2.6 %), A*01:01 ∼ B*57:01 ∼ C*06:02 ∼ DRB1*07:01 ∼ DQB1*03:03 (2.4 %), and A*01:01 ∼ B*37:01 ∼ C*06:02 ∼ DRB1*10:01 ∼ DQB1*05:01 (1.4 %), showing significant inter-regional differences. We observed distinct allele distributions and high haplotype diversity, with the India South UCBB population displaying the highest number of unique haplotypes (63 %). Genetic distance analysis and multidimensional scaling revealed genetic relatedness between the regional and sub-regional populations. These findings offer key insights into Indian HLA diversity, inform donor recruitment strategies, enhance understanding of genetic relatedness to neighbouring populations, and highlight the need for population-specific repositories.
Chronic alloimmune-mediated injury in kidney transplantation continues to elude effective treatment and negatively impact outcomes. With increased utilization of simultaneous liver and kidney transplants (SLK), many have...Chronic alloimmune-mediated injury in kidney transplantation continues to elude effective treatment and negatively impact outcomes. With increased utilization of simultaneous liver and kidney transplants (SLK), many have observed that these recipients have improved kidney graft survival when compared to kidney alone recipients. This intriguing observation has incited a growing group of studies that aim to elucidate the mechanisms behind the liver allograft's ability to modulate recipient's alloimmune responses against the kidney allograft. In this article, we will provide a brief overview of donor specific antibodies (DSA)-mediated injury of kidney allograft, the impact of SLK on recipients with pre-existing DSA, and current theorized pathways through which the liver allograft confers immune tolerance.
The role of anti-HLA antibodies in solid organ transplantation has long been recognized, particularly with respect to graft rejection and patient survival. This study aimed to evaluate the impact of de novo donor-specifi...The role of anti-HLA antibodies in solid organ transplantation has long been recognized, particularly with respect to graft rejection and patient survival. This study aimed to evaluate the impact of de novo donor-specific antibodies (DSA) on graft failure and mortality in Polish lung transplant recipients. The study included 112 patients who had a well-defined immune status prior to transplantation. The presence and specificity of anti-HLA antibodies were assessed using the Luminex platform. We identified 26 patients (23%) as having increased immunological risk. Pre-transplant DSAs were detected in 13 recipients. Post-transplantation, the development of de novo DSAs was evaluated, with a cumulative incidence of 11%. Our findings confirmed that lung transplant recipients with donor-specific immunization frequently develop antibodies targeting antigens at the HLA-DQ locus. Notably, 90% (9 out of 10) of immunized patients exhibited high mean fluorescence intensity (MFI) levels of anti-DQ antibodies. These antibodies were shown to be harmful, with mortality significantly higher in immunized patients: 50% of patients with anti-DQ antibodies died, compared to only 15% in those without DSAs. This is the first study of its kind in the Polish population, confirming that strategies to prevent and treat DQ DSAs may improve outcomes for lung transplant recipients.
Łacina P, Crossland RE, Siemaszko J
… +14 more, Szeremet A, Majcherek M, Czyż A, Sobczyk-Kruszelnicka M, Fidyk W, Solarska I, Nasiłowska-Adamska B, Skowrońska P, Bieniaszewska M, Tomaszewska A, Basak GW, Giebel S, Wróbel T, Bogunia-Kubik K
Cytomegalovirus (CMV) reactivation is a common complication after allogeneic haematopoietic stem cell transplantation (HSCT). It occurs in over a third of alloHSCT recipients, and is a major source of post-transplant mor...Cytomegalovirus (CMV) reactivation is a common complication after allogeneic haematopoietic stem cell transplantation (HSCT). It occurs in over a third of alloHSCT recipients, and is a major source of post-transplant mortality. miRNAs are small non-coding RNA molecules that are important regulators of gene expression. They are often dysregulated during disease, including transplant-related complications. Due to their presence in all body fluids, they can be used as potential biomarkers. In this study, we performed miRNA profiling to find differentially expressed host miRNAs in serum that could be linked to CMV reactivation after HSCT. CMV reactivation was confirmed by detection of CMV DNA in serum. We identified two miRNAs, miR-302d-3p (p < 0.001) and miR-6721-5p (p = 0.018), as differentially expressed between day + 30 and + 90 after HSCT in patients with CMV reactivation. Of these, miR-302d-3p was confirmed as overexpressed on day + 30 in a verification analysis performed on an independent cohort of patients (p = 0.027). Furthermore, we analysed two genetic variants in the gene coding for miR-302d-3p. We found one of them, rs13136737, to be significantly associated with CMV reactivation (p = 0.015). In conclusion, our study showed that miR-302d-3p is dysregulated and that its genetic variant rs13136737 may be important in development of CMV reactivation after HSCT.
INTRODUCTION: Influenza A virus (IAV) and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) coinfection have been shown to intensify symptoms and, therefore, could be considered a significant threat to suscept...INTRODUCTION: Influenza A virus (IAV) and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) coinfection have been shown to intensify symptoms and, therefore, could be considered a significant threat to susceptible populations. Although vaccination is an effective and safe strategy to be used against viruses, large amounts of mutations in these two viruses remain an issue for vaccine design, as some strains of the IAV and SARS-CoV-2 are still present worldwide, regardless of the introduction of various vaccines to protect against the viruses. METHODS: We employed a bioinformatic approach to design a hybrid vaccine based on the epitopes of IAV and SARS-CoV-2, which are potentially conserved against mutations and are safe in terms of toxicity and allergenicity, yet promise to induce long-lasting innate and adaptive immune responses. RESULTS: The selected peptides demonstrated a potentially effective immune response and showed promise as safe and suitable for hybrid vaccine design. We selected epitopes focusing on their binding quality to cytotoxic T lymphocyte (CTL), helper T lymphocyte (HTL), and B cell. For instance, CTL's epitopes were checked using sequence-based prediction methods, and their entropy and antigenicity were confirmed. Similarly, all epitopes were selected based on their ability to induce robust humoral and cell-mediated immunity, respectively. CONCLUSION: Hybrid vaccine design, utilizing our approach, can lead to next-generation hybrid vaccines that help the healthcare system establish a more consistent defense against both viruses for the susceptible population.
Whether hydroxychloroquine intervention can improve pregnancy outcomes in patients with recurrent pregnancy loss patients (RPL) with antinuclear antibodies (ANAs) positive is still unknown. RPL patients who were positive...Whether hydroxychloroquine intervention can improve pregnancy outcomes in patients with recurrent pregnancy loss patients (RPL) with antinuclear antibodies (ANAs) positive is still unknown. RPL patients who were positive for ANAs without rheumatism were recruited from the Reproductive Medicine Center of the Second Affiliated Hospital of Soochow University from January 2016 to December 2023 (N = 104). Normal healthy pregnant women were selected as the Control group (N = 100). ANA-positive pregnant women who had experienced one sporadic abortion were included in the SA group (N = 42). Patients in both RPL and SA groups received 0.2-0.4 g hydroxychloroquine daily. General data and pregnancy outcomes were compared among the three groups. Patients with RPL were divided into three subgroups according to ANA titers: 1:100 (n = 81), 1:320 (n = 18) and 1:1000 (n = 5). The differences in the pregnancy outcomes among the subgroups were compared. We found the abortion rates in RPL and SA groups were greater than that in the control group (P < 0.05). The incidence of pregnancy complications was greater in the RPL group than in the control group (P < 0.05). The full-term live birth rates in RPL and SA groups were lower than that in the control group (P < 0.05). There were significant differences in the abortion rate among patients with RPL with different ANA titers (P < 0.05). We concluded that adverse pregnancy outcomes, such as abortion and complications during pregnancy, are still more common in ANA-positive RPL patients than in normal pregnant women despite treatment. The ANA titer is closely related to the abortion rate among RPL patients during sequent pregnancy.
Studies have demonstrated that the liver acts as an immunologic sink when transplanted with another organ from the same donor. In this work, we review evidence of this phenomenon in kidney-liver and heart-liver transplan...Studies have demonstrated that the liver acts as an immunologic sink when transplanted with another organ from the same donor. In this work, we review evidence of this phenomenon in kidney-liver and heart-liver transplantation. We then explore the pros and cons of this strategy as a way of helping to transplant end stage heart failure patients who are highly allosensitized but do not have severe liver disease. By considering this strategy in the context of alternative therapies, we explain the types of patients who may benefit from it.
PURPOSE: The ABO blood group gene, a protein-coding gene on chromosome 9, is essential for identifying a person's blood group phenotype. A, B, and O are the three primary alleles of the gene. The ABO blood grouping syste...PURPOSE: The ABO blood group gene, a protein-coding gene on chromosome 9, is essential for identifying a person's blood group phenotype. A, B, and O are the three primary alleles of the gene. The ABO blood grouping system is crucial for organ transplantation and blood transfusion because it helps to avoid potentially fatal unfavorable immunological reactions. This study looks into the codon structuring pattern, Codon Usage Bias (CUB) as well as RNA alteration sites in the ABO blood group alleles of Homo sapiens. RESULTS: The ABO blood group gene has high GC content and low CUB, which both point to significant variation in synonymous codon use. It was discovered that 18 codons were underrepresented and 13 codons were overrepresented in ABO gene. The research indicates that the evolution of ABO blood group genes is shaped by two primary factors: mutational force as well as natural selection, which plays a more substantial role (53.20%) compared to mutational force (46.80%). Transition sites were more than transversion sites. RNA editing sites analysis identified the greatest substitution of C by T nucleotides in ABO alleles. CONCLUSION: Overall, the discoveries broaden our understanding of codon utilization mechanisms and the RNA alteration sites in human ABO blood group alleles.
Klebsiella pneumoniae (K. pneumoniae) is a Gram-negative clinically relevant pathogen responsible for causing nosocomial infections. This bacterium is resistant to a spectrum of antibiotics and has been listed under "cri...Klebsiella pneumoniae (K. pneumoniae) is a Gram-negative clinically relevant pathogen responsible for causing nosocomial infections. This bacterium is resistant to a spectrum of antibiotics and has been listed under "critical" priority for research and development of new antibiotics and alternative strategies. Reverse vaccinology was used to construct multiepitope vaccine candidates against six K. pneumoniae outer membrane proteins. Predicted B- and T-cell epitopes were evaluated using various bioinformatic tools to confirm their antigenic and non-allergenic nature. Additionally, the global population coverage of the epitopes was observed to be 75% as per in silico analysis. These epitopes were then used for the design and in silico characterization of seven multiepitope vaccine candidates. Molecular docking and simulation studies demonstrated that 5 out of the 7 vaccine candidates formed strong stable interactions with TLR2 and TLR4. Furthermore, in silico immunization simulations demonstrated the ability of these candidates to elicit effective immune responses. Lastly, high codon adaptation Index (CAI) values ensured that the candidates can be expressed in the Escherichia coli K12 host system following codon optimized multiepitope cloning. Importantly, the high antigenicity and global coverage of the combined construct represents a novel combination not previously reported. Based on our findings, the designed multiepitope vaccine candidates have potential as a vaccination strategy against K. pneumoniae.
Quality Assurance (QA) plays a pivotal role in safeguarding the accuracy and reliability of laboratory test results, which are integral to informed clinical decision-making and optimal patient outcomes. External Quality...Quality Assurance (QA) plays a pivotal role in safeguarding the accuracy and reliability of laboratory test results, which are integral to informed clinical decision-making and optimal patient outcomes. External Quality Assessment (EQA), a cornerstone of QA, is mandated by international standards such as ISO/IEC 15189 to ensure laboratory competence and comparability across institutions. Despite India's position as the third-highest organ transplanting nation globally, a significant void persists in the availability of accredited EQA providers specialising in transplant immunology and immunophenotyping (IPT). Recognising this critical gap, the Chimera Translational Research Fraternity (CTRF) undertook a comprehensive gap analysis in 2019. The majority of participating laboratories articulated a strong demand for a locally accessible, cost-effective EQA program tailored to the specific requirements of the transplant immunology domain. In response, Chimera EQA initiated a structured implementation strategy that included the recruitment of qualified personnel, the formation of a specialised core committee to develop technical protocols, the establishment of a Quality Management System (QMS), conducting pilot surveys and internal audits. These foundational steps preceded the formal application for ISO/IEC 17043 accreditation through the National Accreditation Board for Testing and Calibration Laboratories (NABL), a division of Quality Council of India (QCI). Following the successful closure of nonconformities (NCs) and rigorous assessments, the program obtained ISO/IEC 17043 accreditation (initially ISO/IEC 17043:2010 and now updated to ISO/IEC 17043:2023) covering 25 analytes, officially establishing India's first accredited EQA program in this niche field. Since its inception, Chimera EQA has successfully conducted 82 PT surveys, overcoming formidable challenges such as securing clinical samples, raising awareness, and developing a robust digital infrastructure. This article demonstrates the successful implementation of this program and shows that, even within limited resources, an accredited EQA initiative is feasible. It can contribute to enhancing diagnostic quality, regulatory compliance, and international recognition in transplant immunology and IPT.