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In this study, we investigated the effect of adenosine and its derivative cordycepin on the production yield of a recombinant human monoclonal antibody (adalimumab) in two commonly used Chinese Hamster ovary (CHO) cell lines that have different gene amplification systems, namely CHO-DHFR - and GS-CHO knockout (GS-KO CHO) cells and that were grown in batch culture, with and without glucose feeding. The results showed that adenosine suppressed the cell growth rate and increased the fraction of cells in S phase of the cell cycle for both CHO cell lines. Different concentrations and exposure times of adenosine feeding were tested. The optimal yield of adalimumab production was achieved with the addition of 1 mM adenosine on day 2 after start of the batch culture. Adenosine could significantly improve antibody titers and productivity in both CHO cell lines in cultures without glucose feeding. However, upon glucose feeding, adenosine did not improve antibody titers in CHO-DHFR - cells but extended culture duration and significantly increased antibody titers in GS-KO CHO cells. Therefore, adenosine supplementation might be useful for antibody production in GS-KO CHO cells in medium- to large-scale batches. In case of cordycepin, a derivative of adenosine, CHO-DHFR - cells required higher concentration of cordycepin than GS-KO CHO cells around 10 times to display the changes in cell growth and cell cycle. Moreover, cordycepin could significantly increase antibody titers only in CHO-DHFR - cell cultures without glucose feeding.

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Background and objectives: Use of convalescent plasma for coronavirus disease 2019 (COVID-19) treatment has gained interest worldwide. However, there is lack of evidence on its dosing, safety and effectiveness. Until data from clinical studies are available to provide solid evidence for worldwide applicable guidelines, there is a need to provide guidance to the transfusion community and researchers on this emergent therapeutic option. This paper aims to identify existing key gaps in current knowledge in the clinical application of COVID-19 convalescent plasma (CCP). Material(s) and Method(s): The International Society of Blood Transfusion (ISBT) initiated a multidisciplinary working group with worldwide representation from all six continents with the aim of reviewing existing practices on CCP use from donor, product and patient perspectives. A subgroup of clinical transfusion professionals was formed to draft a document for CCP clinical application to identify the gaps in knowledge in existing literature. Result(s): Gaps in knowledge were identified in the following main domains: study design, patient eligibility, CCP dose, frequency and timing of CCP administration, parameters to assess response to CCP treatment and long-term outcome, adverse events and CCP application in less-resourced countries as well as in paediatrics and neonates. Conclusion(s): This paper outlines a framework of gaps in the knowledge of clinical deployment of CPP that were identified as being most relevant. Studies to address the identified gaps are required to provide better evidence on the effectiveness and safety of CCP use.Copyright © 2020 The Authors. Vox Sanguinis published by John Wiley & Sons Ltd on behalf of International Society of Blood Transfusion

Background and objectives: Use of convalescent plasma for coronavirus disease 2019 (COVID-19) treatment has gained interest worldwide. However, there is lack of evidence on its dosing, safety and effectiveness. Until data from clinical studies are available to provide solid evidence for worldwide applicable guidelines, there is a need to provide guidance to the transfusion community and researchers on this emergent therapeutic option. This paper aims to identify existing key gaps in current knowledge in the clinical application of COVID-19 convalescent plasma (CCP). Material(s) and Method(s): The International Society of Blood Transfusion (ISBT) initiated a multidisciplinary working group with worldwide representation from all six continents with the aim of reviewing existing practices on CCP use from donor, product and patient perspectives. A subgroup of clinical transfusion professionals was formed to draft a document for CCP clinical application to identify the gaps in knowledge in existing literature. Result(s): Gaps in knowledge were identified in the following main domains: study design, patient eligibility, CCP dose, frequency and timing of CCP administration, parameters to assess response to CCP treatment and long-term outcome, adverse events and CCP application in less-resourced countries as well as in paediatrics and neonates. Conclusion(s): This paper outlines a framework of gaps in the knowledge of clinical deployment of CPP that were identified as being most relevant. Studies to address the identified gaps are required to provide better evidence on the effectiveness and safety of CCP use.Copyright © 2020 The Authors. Vox Sanguinis published by John Wiley & Sons Ltd on behalf of International Society of Blood Transfusion

Rabies remains endemic in Zambia. Despite conducting canine vaccinations in Lusaka district, the vaccination coverage and actual seropositivity in the dog population in Lusaka district are rarely evaluated. This study estimated the seropositivity-based immunization coverage in the owned dog population in Lusaka district using the expanded program on immunization cluster survey method. The time-series trend of neutralizing antibodies against rabies in vaccinated dogs was also evaluated. Of 366 dogs in 200 dog-owning households in Lusaka district, blood samples were collected successfully from 251 dogs. In the sampled dogs, 42.2% (106/251) had an antibody titer >= 0.5 IU/mL. When the 115 dogs whose blood was not collected were assumed to be seronegative, the minimum immunization coverage in Lusaka district's owned dog population was estimated at 29.0% (95% confidence interval: 22.4-35.5). It was also found that a single vaccination with certified vaccines is capable of inducing protective levels of antibodies. In contrast, higher antibody titers were observed in multiple-vaccinated dogs than in single-vaccinated dogs, coupled with the observation of a decline in antibody titer over time. These results suggest the importance of continuous booster immunization to maintain herd immunity and provide useful information to plan mass vaccination against rabies in Zambia.

Rabies remains endemic in Zambia. Despite conducting canine vaccinations in Lusaka district, the vaccination coverage and actual seropositivity in the dog population in Lusaka district are rarely evaluated. This study estimated the seropositivity-based immunization coverage in the owned dog population in Lusaka district using the expanded program on immunization cluster survey method. The time-series trend of neutralizing antibodies against rabies in vaccinated dogs was also evaluated. Of 366 dogs in 200 dog-owning households in Lusaka district, blood samples were collected successfully from 251 dogs. In the sampled dogs, 42.2% (106/251) had an antibody titer >= 0.5 IU/mL. When the 115 dogs whose blood was not collected were assumed to be seronegative, the minimum immunization coverage in Lusaka district's owned dog population was estimated at 29.0% (95% confidence interval: 22.4-35.5). It was also found that a single vaccination with certified vaccines is capable of inducing protective levels of antibodies. In contrast, higher antibody titers were observed in multiple-vaccinated dogs than in single-vaccinated dogs, coupled with the observation of a decline in antibody titer over time. These results suggest the importance of continuous booster immunization to maintain herd immunity and provide useful information to plan mass vaccination against rabies in Zambia.

Background & Aims: Helicobacter pylori induces strong inflammatory responses that are directed at clearing the infection, but if not controlled, these responses can be harmful to the host. We investigated the immune-regulatory effects of the innate immune molecule, nucleotide-binding oligomerization domain-like receptors (NLR) family CARD domain-containing 5 (NLRC5), in patients and mice with Helicobacter infection. Method(s): We obtained gastric biopsies from 30 patients in Australia. We performed studies with mice that lack NLRC5 in the myeloid linage (Nlrc5moKO) and mice without Nlrc5 gene disruption (controls). Some mice were gavaged with H pylori SS1 or Helicobacter felis; 3 months later, stomachs, spleens, and sera were collected, along with macrophages derived from bone marrow. Human and mouse gastric tissues and mouse macrophages were analyzed by histology, immunohistochemistry, immunoblots, and quantitative polymerase chain reaction. THP-1 cells (human macrophages, controls) and NLRC5-/- THP-1 cells (generated by CRISPR-Cas9 gene editing) were incubated with Helicobacter and gene expression and production of cytokines were analyzed. Result(s): Levels of NLRC5 messenger RNA were significantly increased in gastric tissues from patients with H pylori infection, compared with patients without infection (P <.01), and correlated with gastritis severity (P <.05). H pylori bacteria induced significantly higher levels of chemokine and cytokine production by NLRC5-/- THP-1 macrophages than by control THP-1 cells (P <.05). After 3 months of infection with H felis, Nlrc5mo-KO mice developed gastric hyperplasia (P <.0001), splenomegaly (P <.0001), and increased serum antibody titers (P <.01), whereas control mice did not. Nlrc5mo-KO mice with chronic H felis infection had increased numbers of gastric B-cell follicles expressing CD19 (P <.0001); these follicles had features of mucosa-associated lymphoid tissue lymphoma. We identified B-cell-activating factor as a protein

A 4-month-old boy of non-consanguineous parents was referred to the Paediatric Endocrinology Department for growth failure complicated by hypoglycaemia. During hypoglycaemia, his GH and IGF-1 remained undetectable, and other causes of growth failure were excluded. He was commenced on recombinant-hGH (rhGH). After an initial good response, growth slowed despite increasing doses of rhGH, up to 7.5 mg/m2/week. A poor IGF-1 response to rhGH was noted. Compliance issues were excluded. GH deficiency was re-confirmed by an Arginine-Glucagon provocation test. The rhGH was discontinued at 4 years 5 months because growth response criteria were not met. The possibility of GH resistance due to autoantibodies to rhGH was confirmed using an I125 competitive binding research assay at SydPath with an extremely high titre of 1:2,560. Family history revealed an older sister who also had documented GH deficiency. She grew well on rhGH for 10 years followed by a slowdown in growth during puberty. She has now been off rhGH for 18 months and has a very low positive GH antibody titre. DNA testing of GH1 gene sequencing in the family is underway.1 Due to the rare combination of severe GH deficiency and high GH antibodies, a trial of recombinant IGF-1 appears to be the only therapeutic option.2 References 1. Argente J, Tatton-Brown K, Lehwalder D, et al. Genetics of growth disorders-which patients require genetic testing? Front Endocrinol 2019; 10: 602. 2. Ranke MB. Treatment with recombinant human insulin-like growth factor-1 in severe primary IGF deficiency and beyond. Horm Res Paediatr 2015; 83: 358-60. Copyright © 2020

Background: Annual influenza vaccination is recommended to all individuals over 6 months of age, including predominantly antibody deficiency (PAD) patients. Vaccination responses are typically evaluated by serology, and because PAD patients are by definition impaired in generating IgG and receive immunoglobulin replacement therapy (IgRT), it remains unclear whether they can mount an antigen-specific response. Objective(s): To quantify and characterise the antigen-specific memory B (Bmem) cell compartment in healthy controls and PAD patients following an influenza booster vaccination. Method(s): Recombinant hemagglutinin (HA) from the A/Michigan/2015 H1N1 (AM15) strain with an AviTag was generated in a mammalian cell line, and following targeted biotinylation, was tetramerised with BUV395 or BUV737 streptavidin conjugates. Multicolour flow cytometry was applied on blood samples before and 28 days after booster influenza vaccination in 16 healthy controls and five PAD patients with circulating Bmem cells. Result(s): Recombinant HA tetramers were specifically recognised by 0.5-1% of B cells in previously vaccinated healthy adults. HA-specific Bmem cell numbers were significantly increased following booster vaccination and predominantly expressed IgG1. Similarly, PAD patients carried HA-specific Bmem cells, predominantly expressing IgG1. However, these numbers were lower than in controls and did not increase following booster vaccination. Conclusion(s): We have successfully identified AM15-specific Bmem cells in healthy controls and PAD patients. The presence of antigen-specific Bmem cells could offer an additional diagnostic tool to aid in the clinical diagnosis of PAD. Furthermore, alterations in the number or immunophenotype of HA-specific Bmem cells post-booster vaccination could assist in the evaluation of immune responses in individuals receiving IgRT.Copyright © 2020 The Authors. Clinical & Translational Immunology published by John Wiley & Sons Australia, Ltd on

Aim: In Australia, the risk of hepatitis B virus (HBV) transmission from single sexual contact is low. This, combined with assumed widespread immunity from vaccination, has resulted in a lack of clarity surrounding the necessity for hepatitis B post-exposure prophylaxis following recent sexual assault. Method(s): This retrospective audit was conducted through the Victorian Forensic Paediatric Medical Service (VFPMS) at the Royal Children's Hospital, Melbourne, Australia. Subjects were patients aged 13-17 years who presented to VFPMS between 1 January 2007 and 31 December 2016 for forensic medical examination following an alleged penetrative sexual assault. Data collected included subject demographics, immunisation status, route of potential HBV exposure, time between alleged sexual assault and presentation, whether HBsAb levels were tested and the results and whether HBV prophylaxis was administered to the subject and its timing. Result(s): A total of 2121 records were reviewed, and 420 subjects were found to be eligible for inclusion; 26.2% (n = 110) had HBsAb levels measured at initial presentation. Of these 110 subjects, 45.5% (n = 50) had titre levels less than 10 (deemed to be non-protective) and were therefore vulnerable to HBV infection. Of the 420 subjects, 4.5% (n = 19) received HBV prophylaxis as a result of their assessment. Conclusion(s): Results suggest that a high proportion of Australian adolescents presenting following recent sexual assault may be at risk of hepatitis B infection. Very few received timely prophylaxis. Follow-up attendance rates were poor. Administration of the hepatitis B booster vaccine at the point of contact may reduce the risk of HBV infection in this group of adolescents.Copyright © 2020 Paediatrics and Child Health Division (The Royal Australasian College of Physicians)

Waldenstrom macroglobulinaemia (WM) is an indolent non-Hodgkin lymphoma which usually presents with symptoms related to infiltration of bone marrow or other tissues like lymph nodes, liver or spleen and has certain unusual clinical manifestations, e.g., renal and central nervous system (CNS) involvement. It also has an array of laboratory features including hypersecretion of IgM, cryoglobulinaemia, increased plasma viscosity and identification of mutated MYD88L265P in more than 90% of cases. In this review, we aim to provide a guide to the laboratory investigations recommended for WM at initial diagnosis and at follow-up. A discussion on the nuances of diagnosis and differential diagnoses is followed by bone marrow (BM) assessment, measurement of paraprotein and other ancillary investigations. Recommendations are provided on laboratory work-up at diagnosis, in the asymptomatic follow-up phase, and during and post-treatment. Finally, we briefly discuss the implications of laboratory diagnosis in regard to recruitment and monitoring on clinical trials.Copyright © 2019

Background & Aims: Helicobacter pylori induces strong inflammatory responses that are directed at clearing the infection, but if not controlled, these responses can be harmful to the host. We investigated the immune-regulatory effects of the innate immune molecule, nucleotide-binding oligomerization domain-like receptors (NLR) family CARD domain-containing 5 (NLRC5), in patients and mice with Helicobacter infection. Method(s): We obtained gastric biopsies from 30 patients in Australia. We performed studies with mice that lack NLRC5 in the myeloid linage (Nlrc5moKO) and mice without Nlrc5 gene disruption (controls). Some mice were gavaged with H pylori SS1 or Helicobacter felis; 3 months later, stomachs, spleens, and sera were collected, along with macrophages derived from bone marrow. Human and mouse gastric tissues and mouse macrophages were analyzed by histology, immunohistochemistry, immunoblots, and quantitative polymerase chain reaction. THP-1 cells (human macrophages, controls) and NLRC5-/- THP-1 cells (generated by CRISPR-Cas9 gene editing) were incubated with Helicobacter and gene expression and production of cytokines were analyzed. Result(s): Levels of NLRC5 messenger RNA were significantly increased in gastric tissues from patients with H pylori infection, compared with patients without infection (P <.01), and correlated with gastritis severity (P <.05). H pylori bacteria induced significantly higher levels of chemokine and cytokine production by NLRC5-/- THP-1 macrophages than by control THP-1 cells (P <.05). After 3 months of infection with H felis, Nlrc5mo-KO mice developed gastric hyperplasia (P <.0001), splenomegaly (P <.0001), and increased serum antibody titers (P <.01), whereas control mice did not. Nlrc5mo-KO mice with chronic H felis infection had increased numbers of gastric B-cell follicles expressing CD19 (P <.0001); these follicles had features of mucosa-associated lymphoid tissue lymphoma. We identified B-cell-activating factor as a protein

A 4-month-old boy of non-consanguineous parents was referred to the Paediatric Endocrinology Department for growth failure complicated by hypoglycaemia. During hypoglycaemia, his GH and IGF-1 remained undetectable, and other causes of growth failure were excluded. He was commenced on recombinant-hGH (rhGH). After an initial good response, growth slowed despite increasing doses of rhGH, up to 7.5 mg/m2/week. A poor IGF-1 response to rhGH was noted. Compliance issues were excluded. GH deficiency was re-confirmed by an Arginine-Glucagon provocation test. The rhGH was discontinued at 4 years 5 months because growth response criteria were not met. The possibility of GH resistance due to autoantibodies to rhGH was confirmed using an I125 competitive binding research assay at SydPath with an extremely high titre of 1:2,560. Family history revealed an older sister who also had documented GH deficiency. She grew well on rhGH for 10 years followed by a slowdown in growth during puberty. She has now been off rhGH for 18 months and has a very low positive GH antibody titre. DNA testing of GH1 gene sequencing in the family is underway.1 Due to the rare combination of severe GH deficiency and high GH antibodies, a trial of recombinant IGF-1 appears to be the only therapeutic option.2 References 1. Argente J, Tatton-Brown K, Lehwalder D, et al. Genetics of growth disorders-which patients require genetic testing? Front Endocrinol 2019; 10: 602. 2. Ranke MB. Treatment with recombinant human insulin-like growth factor-1 in severe primary IGF deficiency and beyond. Horm Res Paediatr 2015; 83: 358-60. Copyright © 2020

Background: Annual influenza vaccination is recommended to all individuals over 6 months of age, including predominantly antibody deficiency (PAD) patients. Vaccination responses are typically evaluated by serology, and because PAD patients are by definition impaired in generating IgG and receive immunoglobulin replacement therapy (IgRT), it remains unclear whether they can mount an antigen-specific response. Objective(s): To quantify and characterise the antigen-specific memory B (Bmem) cell compartment in healthy controls and PAD patients following an influenza booster vaccination. Method(s): Recombinant hemagglutinin (HA) from the A/Michigan/2015 H1N1 (AM15) strain with an AviTag was generated in a mammalian cell line, and following targeted biotinylation, was tetramerised with BUV395 or BUV737 streptavidin conjugates. Multicolour flow cytometry was applied on blood samples before and 28 days after booster influenza vaccination in 16 healthy controls and five PAD patients with circulating Bmem cells. Result(s): Recombinant HA tetramers were specifically recognised by 0.5-1% of B cells in previously vaccinated healthy adults. HA-specific Bmem cell numbers were significantly increased following booster vaccination and predominantly expressed IgG1. Similarly, PAD patients carried HA-specific Bmem cells, predominantly expressing IgG1. However, these numbers were lower than in controls and did not increase following booster vaccination. Conclusion(s): We have successfully identified AM15-specific Bmem cells in healthy controls and PAD patients. The presence of antigen-specific Bmem cells could offer an additional diagnostic tool to aid in the clinical diagnosis of PAD. Furthermore, alterations in the number or immunophenotype of HA-specific Bmem cells post-booster vaccination could assist in the evaluation of immune responses in individuals receiving IgRT.Copyright © 2020 The Authors. Clinical & Translational Immunology published by John Wiley & Sons Australia, Ltd on

Aim: In Australia, the risk of hepatitis B virus (HBV) transmission from single sexual contact is low. This, combined with assumed widespread immunity from vaccination, has resulted in a lack of clarity surrounding the necessity for hepatitis B post-exposure prophylaxis following recent sexual assault. Method(s): This retrospective audit was conducted through the Victorian Forensic Paediatric Medical Service (VFPMS) at the Royal Children's Hospital, Melbourne, Australia. Subjects were patients aged 13-17 years who presented to VFPMS between 1 January 2007 and 31 December 2016 for forensic medical examination following an alleged penetrative sexual assault. Data collected included subject demographics, immunisation status, route of potential HBV exposure, time between alleged sexual assault and presentation, whether HBsAb levels were tested and the results and whether HBV prophylaxis was administered to the subject and its timing. Result(s): A total of 2121 records were reviewed, and 420 subjects were found to be eligible for inclusion; 26.2% (n = 110) had HBsAb levels measured at initial presentation. Of these 110 subjects, 45.5% (n = 50) had titre levels less than 10 (deemed to be non-protective) and were therefore vulnerable to HBV infection. Of the 420 subjects, 4.5% (n = 19) received HBV prophylaxis as a result of their assessment. Conclusion(s): Results suggest that a high proportion of Australian adolescents presenting following recent sexual assault may be at risk of hepatitis B infection. Very few received timely prophylaxis. Follow-up attendance rates were poor. Administration of the hepatitis B booster vaccine at the point of contact may reduce the risk of HBV infection in this group of adolescents.Copyright © 2020 Paediatrics and Child Health Division (The Royal Australasian College of Physicians)

Waldenstrom macroglobulinaemia (WM) is an indolent non-Hodgkin lymphoma which usually presents with symptoms related to infiltration of bone marrow or other tissues like lymph nodes, liver or spleen and has certain unusual clinical manifestations, e.g., renal and central nervous system (CNS) involvement. It also has an array of laboratory features including hypersecretion of IgM, cryoglobulinaemia, increased plasma viscosity and identification of mutated MYD88L265P in more than 90% of cases. In this review, we aim to provide a guide to the laboratory investigations recommended for WM at initial diagnosis and at follow-up. A discussion on the nuances of diagnosis and differential diagnoses is followed by bone marrow (BM) assessment, measurement of paraprotein and other ancillary investigations. Recommendations are provided on laboratory work-up at diagnosis, in the asymptomatic follow-up phase, and during and post-treatment. Finally, we briefly discuss the implications of laboratory diagnosis in regard to recruitment and monitoring on clinical trials.Copyright © 2019

Purpose: The aim of this study was to determine if measurement of B cell protective immunity was associated with susceptibility to sinopulmonary infection in kidney transplant recipients. Methods and Materials: A prospective cohort of 168 patients with stable graft function (median 4.1 years) underwent assessment of B-lymphocyte antigen CD19 (CD19+) cell number, immunoglobulin G concentration, and seroresponses to influenza vaccination upon study entry. Patients received a single dose of a trivalent, seasonal influenza vaccine. Result(s): After 2 years follow-up, 31 patients (18%) developed sinopulmonary infection. CD19+ cell number was strongly associated with future sinopulmonary infection. A higher proportion of patients with CD19+ cell counts below the fifth percentile for controls developed sinopulmonary infections than those above the fifth percentile, 30% (23 of 77 patients) compared with 9% (7 of 79 patients; P =.001). There was a trend toward a higher proportion of patients with reduced immunoglobulin G concentrations developing infections than in the normal range for controls, 29% (14 of 48 patients) compared with 15% (16 of 108 patients; P =.060). Influenza vaccination seroresponses were poor in patients and controls such that they could not be used to identify a subgroup of patients at high risk for the development of severe pulmonary infection. Conclusion(s): Monitoring B-cell numbers represents a simple, inexpensive means of stratifying transplant recipients' risk of sinopulmonary infection.Copyright © 2018 Elsevier Inc.

Introduction: Vaccination is used to generate immune memory, including antibody production against potentially harmful pathogens. Transplant recipients are immunosuppressed to prevent similar processes taking place against alloantigens. Modern immunosuppression significantly reduces seroresponses to vaccination. We hypothesized that seroconversion to vaccination in renal transplant recipients could serve as a functional measure of an individual's immunosuppression adequacy by predicting future rejection episodes while a lack of seroconversion might predict subsequent infection. Method(s): We prospectively followed a cohort of 151 patients who received the 2009 monovalent pandemic H1N1 vaccine at our institution, until December 2016. Patients were considered seroconverters (n=48) or non-seroconverters (n=103) with seroconversion defined as a >=4-fold increase in antibody titre from pre-vaccination levels. Time to first rejection or serious infection (infection requiring hospitalisation) was compared between seroconverters and non-seroconverters using Cox regression. Additionally, whether prior rejection or serious infection predicted seroconversion was assessed by logistic regression. Regression models were adjusted for age, sex, transplant duration, peak PRA, eGFR and mycophenolate dose. eGFR, patient and graft survival were also examined. Data were obtained from the Australia and New Zealand Dialysis and Transplant Registry (ANZDATA) and medical record review. Result(s): The included patients had a mean age of 51.2+/-12.6 years, 55.6% were male, 91% had first transplants and the median time post-transplant was 3.7 years (range 0.1-27.9 years). Immunosuppression consisted of a calcineurin inhibitor in 90%, mycophenolate in 81.5% and prednisolone in 63.6%. Twenty-six rejection episodes occurred in the follow up period. Seroconversion was not associated with an increased risk of rejection on univariable or multivariable analysis (HR 1.20, 95% CI 0.50-2.83, p=0.678). F

Purpose: Vaccination generates immune memory, including antibodies against harmful pathogens. Transplant immunosuppression prevents similar processes occurring against alloantigens. Transplant recipients have signifcantly reduced seroresponses to vaccination. We hypothesized that in renal transplant recipients, seroconversion to vaccination may predict future rejection episodes while a lack of seroconversion might predict subsequent infection. Method(s): We followed 151 patients who received the 2009 monovalent pandemic H1N1 vaccine at our institution, until December 2016. Patients were considered seroconverters (n=48) or non-seroconverters (n=103) (seroconversion defned as a >=4-fold increase in antibody titre). Time to frst rejection or serious infection (infection requiring hospitalisation) was compared between seroconverters and non-seroconverters using Cox regression. In addition, prior rejection or serious infection were assessed by logistic regression as predictors of seroconversion. Regression models were adjusted for age, sex, transplant duration, peak PRA, eGFR and mycophenolate dose. eGFR, patient and graft survival were also examined. Registry (ANZDATA) and medical record data was used. Result(s): The cohort had a mean age of 51.2+/-12.6 years, 55.6% were male, 91% had frst transplants. The median time post-transplant was 3.7 years (range 0.1-27.9 years). Immunosuppression consisted of a calcineurin inhibitor in 90%, mycophenolate in 81.5% and prednisolone in 63.6%. Twenty-six rejection episodes occurred in the follow up period. Seroconversion was not associated with an increased risk of rejection on univariable or multivariable analysis (HR 1.20, 95% CI 0.50-2.83, p=0.678). Fifty-eight patients developed a serious infection in the follow-up period. Seroconversion was not associated with subsequent serious infection on univariable or multivariable analysis (HR 0.94 95% CI 0.50-1.75, p=0.835). Seroconversion did not predict patient or graft survival and u

Purpose: The aim of this study was to determine if measurement of B cell protective immunity was associated with susceptibility to sinopulmonary infection in kidney transplant recipients. Methods and Materials: A prospective cohort of 168 patients with stable graft function (median 4.1 years) underwent assessment of B-lymphocyte antigen CD19 (CD19+) cell number, immunoglobulin G concentration, and seroresponses to influenza vaccination upon study entry. Patients received a single dose of a trivalent, seasonal influenza vaccine. Result(s): After 2 years follow-up, 31 patients (18%) developed sinopulmonary infection. CD19+ cell number was strongly associated with future sinopulmonary infection. A higher proportion of patients with CD19+ cell counts below the fifth percentile for controls developed sinopulmonary infections than those above the fifth percentile, 30% (23 of 77 patients) compared with 9% (7 of 79 patients; P =.001). There was a trend toward a higher proportion of patients with reduced immunoglobulin G concentrations developing infections than in the normal range for controls, 29% (14 of 48 patients) compared with 15% (16 of 108 patients; P =.060). Influenza vaccination seroresponses were poor in patients and controls such that they could not be used to identify a subgroup of patients at high risk for the development of severe pulmonary infection. Conclusion(s): Monitoring B-cell numbers represents a simple, inexpensive means of stratifying transplant recipients' risk of sinopulmonary infection.Copyright © 2018 Elsevier Inc.

Anti-myeloperoxidase nephritogenic autoimmunity induces severe glomerulonephritis. To assess the therapeutic potential of monoclonal antibodies targeting T helper (Th) subset differentiation determining cytokines, we studied a murine model of anti-myeloperoxidase glomerulonephritis. The temporal participation of T helper subsets was determined by quantitating gene expression of CD4+ T-cells isolated from nephritic kidneys and cytokine production by lymphocytes from nodes draining myeloperoxidase immunization sites. Th17 cytokines (IL-17A and IL-6) rose rapidly but declined as autoimmunity matured when Th1 cytokines (IL-12 and TNF) predominated. Therefore, T helper subset participation in anti-myeloperoxidase autoimmunity is biphasic, with Th17 early and Th1 late. To confirm the functional relevance of this biphasic pattern, we compared systemic anti-myeloperoxidase autoimmunity in wild type, Th17 deficient and Th1 deficient mice. Early, Th1 deficient mice developed similar autoimmunity and glomerulonephritis to wild type mice. However, Th17 deficient mice had significantly reduced anti-myeloperoxidase autoimmunity. In late autoimmunity, Th1 deficient mice developed reduced autoimmunity and were protected from anti-myeloperoxidase glomerulonephritis. The therapeutic potential of these findings were demonstrated by neutralizing monoclonal antibodies. Targeting IL-23p19 attenuated early Th17 dominated anti-myeloperoxidase autoimmunity and glomerulonephritis but not late phase disease. Targeting IL-12p35 attenuated late phase Th1 dominated anti-myeloperoxidase autoimmunity and glomerulonephritis but not early autoimmunity or glomerulonephritis. Targeting both T helper subsets with an anti-IL-12p40 monoclonal antibody was effective during both early and late phases of anti-myeloperoxidase glomerulonephritis. Thus, definition of dominant T helper differentiating subsets in anti-myeloperoxidase glomerulonephritis by renal CD4+ T-cell cytokine gene expression allows effecti

Introduction: Vaccination is used to generate immune memory, including antibody production against potentially harmful pathogens. Transplant recipients are immunosuppressed to prevent similar processes taking place against alloantigens. Modern immunosuppression significantly reduces seroresponses to vaccination. We hypothesized that seroconversion to vaccination in renal transplant recipients could serve as a functional measure of an individual's immunosuppression adequacy by predicting future rejection episodes while a lack of seroconversion might predict subsequent infection. Method(s): We prospectively followed a cohort of 151 patients who received the 2009 monovalent pandemic H1N1 vaccine at our institution, until December 2016. Patients were considered seroconverters (n=48) or non-seroconverters (n=103) with seroconversion defined as a >=4-fold increase in antibody titre from pre-vaccination levels. Time to first rejection or serious infection (infection requiring hospitalisation) was compared between seroconverters and non-seroconverters using Cox regression. Additionally, whether prior rejection or serious infection predicted seroconversion was assessed by logistic regression. Regression models were adjusted for age, sex, transplant duration, peak PRA, eGFR and mycophenolate dose. eGFR, patient and graft survival were also examined. Data were obtained from the Australia and New Zealand Dialysis and Transplant Registry (ANZDATA) and medical record review. Result(s): The included patients had a mean age of 51.2+/-12.6 years, 55.6% were male, 91% had first transplants and the median time post-transplant was 3.7 years (range 0.1-27.9 years). Immunosuppression consisted of a calcineurin inhibitor in 90%, mycophenolate in 81.5% and prednisolone in 63.6%. Twenty-six rejection episodes occurred in the follow up period. Seroconversion was not associated with an increased risk of rejection on univariable or multivariable analysis (HR 1.20, 95% CI 0.50-2.83, p=0.678). F

Objective: To perform a systematic review of the current scientific literature in order to identify variables associated with patient prognosis in autoimmune encephalitis. Method(s): We performed a systematic literature search using MEDLINE, Embase, PubMed and PsychInfo databases. We selected studies that explored the correlation between early clinical and paraclinical findings, and patient outcomes. Data was extracted, analyzed and recorded in accordance with the Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) guidelines. Result(s): Forty four publications detailing 2823 subjects matched our inclusion criteria. There was considerable heterogeneity in methodology, patient profile, investigation results and clinical outcome measures. Findings were often discrepant for cases of anti-NMDAR encephalitis when compared with other causes of autoimmune encephalitis. Delay in immunotherapy contributed to a variety of worse outcomes for patients with different subsets of autoimmune encephalitis. Altered consciousness, ICU admission and no use of immunotherapy were variables associated with poor prognosis in anti-NMDAR encephalitis. Older age, sex, the presence of status epilepticus, CSF abnormalities and MRI changes were unlikely to have significant prognostic value. The influence of antibody titers, autonomic dysfunction and underlying malignancy was unclear. Conclusion(s): A number of variables were identified to have potential predictive value for outcomes in autoimmune encephalitis. Heterogeneous study design, size and quality were major limiting factors in this review.Copyright © 2018 Elsevier Ltd

Purpose: Vaccination generates immune memory, including antibodies against harmful pathogens. Transplant immunosuppression prevents similar processes occurring against alloantigens. Transplant recipients have signifcantly reduced seroresponses to vaccination. We hypothesized that in renal transplant recipients, seroconversion to vaccination may predict future rejection episodes while a lack of seroconversion might predict subsequent infection. Method(s): We followed 151 patients who received the 2009 monovalent pandemic H1N1 vaccine at our institution, until December 2016. Patients were considered seroconverters (n=48) or non-seroconverters (n=103) (seroconversion defned as a >=4-fold increase in antibody titre). Time to frst rejection or serious infection (infection requiring hospitalisation) was compared between seroconverters and non-seroconverters using Cox regression. In addition, prior rejection or serious infection were assessed by logistic regression as predictors of seroconversion. Regression models were adjusted for age, sex, transplant duration, peak PRA, eGFR and mycophenolate dose. eGFR, patient and graft survival were also examined. Registry (ANZDATA) and medical record data was used. Result(s): The cohort had a mean age of 51.2+/-12.6 years, 55.6% were male, 91% had frst transplants. The median time post-transplant was 3.7 years (range 0.1-27.9 years). Immunosuppression consisted of a calcineurin inhibitor in 90%, mycophenolate in 81.5% and prednisolone in 63.6%. Twenty-six rejection episodes occurred in the follow up period. Seroconversion was not associated with an increased risk of rejection on univariable or multivariable analysis (HR 1.20, 95% CI 0.50-2.83, p=0.678). Fifty-eight patients developed a serious infection in the follow-up period. Seroconversion was not associated with subsequent serious infection on univariable or multivariable analysis (HR 0.94 95% CI 0.50-1.75, p=0.835). Seroconversion did not predict patient or graft survival and u

Introduction Animal models have been significant contributors to our understanding of the pathogenesis of many autoimmune diseases, and in some cases have also been useful as pre-clinical models to inform the development of new treatments. A number of animal models have been developed to help understand the pathogenesis of antineutrophil antibody (ANCA)-associated vasculitis (AAV). This commentary will focus on some of the models of myeloperoxidase (MPO)-ANCA associated glomerulonephritis, briefly outlining some of the currently available models (several recent detailed reviews exist) and reviewing some of the recent insights gained from these models in the last two years. Models of MPO-ANCA Associated Glomerulonephritis Models induced by MPO-ANCA-Like Antibodies Transfer of MPO-ANCA like antibodies induced by immunising Mpo-/-mice with native mouse MPO induces proteinuria, haematuria and focal necrotising and crescentic glomerulonephritis [1, 2]. Neutrophil accumulation, assessed several hours after anti-MPO antibody transfer can be induced by low doses of anti-MPO antibodies, but higher doses of anti-MPO antibodies are required for more significant injury [3, 4] and the severity of injury can be increased by pre-treating mice with LPS [5]. The degree of injury, usually measured at day six, is variable, probably due at least in part to the titre and pathogenicity of the antiMPO antibodies raised in the Mpo-/-mice [1, 3, 6]. However, recent publications point towards the capacity for neutrophils to be activated and the numbers of neutrophils in the peripheral blood as important. Xiao et al. found that the 129S6/SvEv and CAST/EiJ mouse strains were markedly more susceptible in this model [7]. Genetic analyses were consistent with the involvement of multiple genetic interactions and studies on neutrophil activation showed that 126S6 neutrophils were more strongly activated by TNF and anti-MPO antibodies [7]. A different approach was taken by Feeley et al [8]. By pre-t

Fibrous dysplasia (FD) is an uncommon benign skeletal disorder, characterized by bone pain, deformities, and the development of pathological fractures. It is caused by osteoblastic lineage differentiation defects, leading to the replacement of normal bone with benign disorganized fibrous connective tissue. Avascular necrosis (AVN) of the femoral head is an insidious condition that can often be challenging to diagnose in its early stages. The pathogenesis of AVN is not well understood; however, it causes femoral head ischemia and collapse, often requiring hip arthroplasty. We report the first case of FD and bilateral AVN of the femoral head in the absence of an antecedent fracture. We postulate several mechanisms to explain how FD may result in AVN; however, further research is required to understand its pathophysiology and thus to guide clinical practice. © 2017 The Authors. JBMR Plus is published by Wiley Periodicals, Inc. on behalf of the American Society for Bone and Mineral Research.

Introduction: We examined the clinical relevance of urinary concentrations of B-cell-activating factor of the tumour necrosis factor family (BAFF) and a proliferation-inducing ligand (APRIL) in systemic lupus erythematosus (SLE). Method(s): We quantified urinary BAFF (uBAFF) by enzyme-linked immunosorbent assay in 85 SLE, 28 primary Sjogren syndrome (pSS), 40 immunoglobulin A nephropathy (IgAN) patients and 36 healthy controls (HCs). Urinary APRIL (uAPRIL) and monocyte chemoattractant protein 1 (uMCP-1) were also quantified. Overall and renal SLE disease activity were assessed using the Systemic Lupus Erythematosus Disease Activity Index 2000. Result(s): uBAFF was detected in 12% (10/85) of SLE patients, but was undetectable in HCs, IgAN and pSS patients. uBAFF was detectable in 28% (5/18) of SLE patients with active nephritis vs 5/67 (7%) of those without (p = 0.03), and uBAFF was significantly higher in active renal patients (p = 0.02) and more likely to be detected in patients with persistently active renal disease. In comparison, uAPRIL and uMCP-1 were detected in 32% (25/77) and 46% (22/48) of SLE patients, respectively. While no difference in proportion of samples with detectable uAPRIL was observed between SLE, HCs and IgAN patients, both uAPRIL and uMCP-1 were significantly detectable in higher proportions of patients with active renal disease. Conclusion(s): uBAFF was detectable in a small but a significant proportion of SLE patients but not in other groups tested, and was higher in SLE patients with active renal disease.Copyright © The Author(s) 2018.

Background: Predominantly antibody deficiencies (PADs) are the most common type of primary immunodeficiency in adults. PADs frequently pass undetected leading to delayed diagnosis, delayed treatment, and the potential for end-organ damage including bronchiectasis. In addition, PADs are frequently accompanied by comorbid autoimmune disease, and an increased risk of malignancy. Objective(s): To characterize the diagnostic and clinical features of adult PAD patients in Victoria, Australia. Method(s): We identified adult patients receiving, or having previously received immunoglobulin replacement therapy for a PAD at four hospitals in metropolitan Melbourne, and retrospectively characterized their clinical and diagnostic features. Result(s): 179 patients from The Royal Melbourne, Alfred and Austin Hospitals, and Monash Medical Centre were included in the study with a median age of 49.7 years (range: 16-87 years), of whom 98 (54.7%) were female. The majority of patients (116; 64.8%) met diagnostic criteria for common variable immunodeficiency (CVID), and 21 (11.7%) were diagnosed with X-linked agammaglobulinemia (XLA). Unclassified hypogammaglobulinemia (HGG) was described in 22 patients (12.3%), IgG subclass deficiency (IGSCD) in 12 (6.7%), and specific antibody deficiency (SpAD) in 4 individuals (2.2%). The remaining four patients had a diagnosis of Good syndrome (thymoma with immunodeficiency). There was no significant difference between the age at diagnosis of the disorders, with the exception of XLA, with a median age at diagnosis of less than 1 year. The median age of reported symptom onset was 20 years for those with a diagnosis of CVID, with a median age at diagnosis of 35 years. CVID patients experienced significantly more non-infectious complications, such as autoimmune cytopenias and lymphoproliferative disease, than the other antibody deficiency disorders. The presence of non-infectious complications was associated with significantly reduced survival in the c

Background: Conjugated pneumococcal vaccine is recommended for kidney transplant recipients, however, their immunogenicity and potential to trigger allograft rejection though generation of de novo anti-human leukocyte antigen antibodies has not been well studied. Method(s): Clinically stable kidney transplant recipients participated in a prospective cohort study and received a single dose of 13-valent conjugate pneumococcal vaccine. Anti-pneumococcal IgG was measured for the 13 vaccine serotypes pre and post vaccination and functional anti-pneumococcal IgG for 4 serotypes post vaccination. Anti-human leukocyte antigen antibodies antibodies were measured before and after vaccination. Kidney transplant recipients were followed clinically for 12 months for episodes of allograft rejection or invasive pneumococcal disease. Result(s): Forty-five kidney transplant recipients participated. Median days between pre and post vaccination serology was 27 (range 21-59). Post vaccination, there was a median 1.1 to 1.7-fold increase in anti-pneumococcal IgG antibody concentrations for all 13 serotypes. Kidney transplant recipients displayed a functional antibody titer >=1:8 for a median of 3 of the 4 serotypes. Post vaccination, there were no de novo anti-human leukocyte antigen antibodies, no episodes of biopsy proven rejection or invasive pneumococcal disease. Conclusion(s): A single dose of 13-valent conjugate pneumococcal vaccine elicits increased titers and breadth of functional anti-pneumococcal antibodies in kidney transplant recipients without stimulating rejection or donor-specific antibodies.Copyright © 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd

Fibrous dysplasia (FD) is an uncommon benign skeletal disorder, characterized by bone pain, deformities, and the development of pathological fractures. It is caused by osteoblastic lineage differentiation defects, leading to the replacement of normal bone with benign disorganized fibrous connective tissue. Avascular necrosis (AVN) of the femoral head is an insidious condition that can often be challenging to diagnose in its early stages. The pathogenesis of AVN is not well understood; however, it causes femoral head ischemia and collapse, often requiring hip arthroplasty. We report the first case of FD and bilateral AVN of the femoral head in the absence of an antecedent fracture. We postulate several mechanisms to explain how FD may result in AVN; however, further research is required to understand its pathophysiology and thus to guide clinical practice. © 2017 The Authors. JBMR Plus is published by Wiley Periodicals, Inc. on behalf of the American Society for Bone and Mineral Research.

Introduction: We examined the clinical relevance of urinary concentrations of B-cell-activating factor of the tumour necrosis factor family (BAFF) and a proliferation-inducing ligand (APRIL) in systemic lupus erythematosus (SLE). Method(s): We quantified urinary BAFF (uBAFF) by enzyme-linked immunosorbent assay in 85 SLE, 28 primary Sjogren syndrome (pSS), 40 immunoglobulin A nephropathy (IgAN) patients and 36 healthy controls (HCs). Urinary APRIL (uAPRIL) and monocyte chemoattractant protein 1 (uMCP-1) were also quantified. Overall and renal SLE disease activity were assessed using the Systemic Lupus Erythematosus Disease Activity Index 2000. Result(s): uBAFF was detected in 12% (10/85) of SLE patients, but was undetectable in HCs, IgAN and pSS patients. uBAFF was detectable in 28% (5/18) of SLE patients with active nephritis vs 5/67 (7%) of those without (p = 0.03), and uBAFF was significantly higher in active renal patients (p = 0.02) and more likely to be detected in patients with persistently active renal disease. In comparison, uAPRIL and uMCP-1 were detected in 32% (25/77) and 46% (22/48) of SLE patients, respectively. While no difference in proportion of samples with detectable uAPRIL was observed between SLE, HCs and IgAN patients, both uAPRIL and uMCP-1 were significantly detectable in higher proportions of patients with active renal disease. Conclusion(s): uBAFF was detectable in a small but a significant proportion of SLE patients but not in other groups tested, and was higher in SLE patients with active renal disease.Copyright © The Author(s) 2018.

Introduction Animal models have been significant contributors to our understanding of the pathogenesis of many autoimmune diseases, and in some cases have also been useful as pre-clinical models to inform the development of new treatments. A number of animal models have been developed to help understand the pathogenesis of antineutrophil antibody (ANCA)-associated vasculitis (AAV). This commentary will focus on some of the models of myeloperoxidase (MPO)-ANCA associated glomerulonephritis, briefly outlining some of the currently available models (several recent detailed reviews exist) and reviewing some of the recent insights gained from these models in the last two years. Models of MPO-ANCA Associated Glomerulonephritis Models induced by MPO-ANCA-Like Antibodies Transfer of MPO-ANCA like antibodies induced by immunising Mpo-/-mice with native mouse MPO induces proteinuria, haematuria and focal necrotising and crescentic glomerulonephritis [1, 2]. Neutrophil accumulation, assessed several hours after anti-MPO antibody transfer can be induced by low doses of anti-MPO antibodies, but higher doses of anti-MPO antibodies are required for more significant injury [3, 4] and the severity of injury can be increased by pre-treating mice with LPS [5]. The degree of injury, usually measured at day six, is variable, probably due at least in part to the titre and pathogenicity of the antiMPO antibodies raised in the Mpo-/-mice [1, 3, 6]. However, recent publications point towards the capacity for neutrophils to be activated and the numbers of neutrophils in the peripheral blood as important. Xiao et al. found that the 129S6/SvEv and CAST/EiJ mouse strains were markedly more susceptible in this model [7]. Genetic analyses were consistent with the involvement of multiple genetic interactions and studies on neutrophil activation showed that 126S6 neutrophils were more strongly activated by TNF and anti-MPO antibodies [7]. A different approach was taken by Feeley et al [8]. By pre-t

Background: Predominantly antibody deficiencies (PADs) are the most common type of primary immunodeficiency in adults. PADs frequently pass undetected leading to delayed diagnosis, delayed treatment, and the potential for end-organ damage including bronchiectasis. In addition, PADs are frequently accompanied by comorbid autoimmune disease, and an increased risk of malignancy. Objective(s): To characterize the diagnostic and clinical features of adult PAD patients in Victoria, Australia. Method(s): We identified adult patients receiving, or having previously received immunoglobulin replacement therapy for a PAD at four hospitals in metropolitan Melbourne, and retrospectively characterized their clinical and diagnostic features. Result(s): 179 patients from The Royal Melbourne, Alfred and Austin Hospitals, and Monash Medical Centre were included in the study with a median age of 49.7 years (range: 16-87 years), of whom 98 (54.7%) were female. The majority of patients (116; 64.8%) met diagnostic criteria for common variable immunodeficiency (CVID), and 21 (11.7%) were diagnosed with X-linked agammaglobulinemia (XLA). Unclassified hypogammaglobulinemia (HGG) was described in 22 patients (12.3%), IgG subclass deficiency (IGSCD) in 12 (6.7%), and specific antibody deficiency (SpAD) in 4 individuals (2.2%). The remaining four patients had a diagnosis of Good syndrome (thymoma with immunodeficiency). There was no significant difference between the age at diagnosis of the disorders, with the exception of XLA, with a median age at diagnosis of less than 1 year. The median age of reported symptom onset was 20 years for those with a diagnosis of CVID, with a median age at diagnosis of 35 years. CVID patients experienced significantly more non-infectious complications, such as autoimmune cytopenias and lymphoproliferative disease, than the other antibody deficiency disorders. The presence of non-infectious complications was associated with significantly reduced survival in the c

Background: Conjugated pneumococcal vaccine is recommended for kidney transplant recipients, however, their immunogenicity and potential to trigger allograft rejection though generation of de novo anti-human leukocyte antigen antibodies has not been well studied. Method(s): Clinically stable kidney transplant recipients participated in a prospective cohort study and received a single dose of 13-valent conjugate pneumococcal vaccine. Anti-pneumococcal IgG was measured for the 13 vaccine serotypes pre and post vaccination and functional anti-pneumococcal IgG for 4 serotypes post vaccination. Anti-human leukocyte antigen antibodies antibodies were measured before and after vaccination. Kidney transplant recipients were followed clinically for 12 months for episodes of allograft rejection or invasive pneumococcal disease. Result(s): Forty-five kidney transplant recipients participated. Median days between pre and post vaccination serology was 27 (range 21-59). Post vaccination, there was a median 1.1 to 1.7-fold increase in anti-pneumococcal IgG antibody concentrations for all 13 serotypes. Kidney transplant recipients displayed a functional antibody titer >=1:8 for a median of 3 of the 4 serotypes. Post vaccination, there were no de novo anti-human leukocyte antigen antibodies, no episodes of biopsy proven rejection or invasive pneumococcal disease. Conclusion(s): A single dose of 13-valent conjugate pneumococcal vaccine elicits increased titers and breadth of functional anti-pneumococcal antibodies in kidney transplant recipients without stimulating rejection or donor-specific antibodies.Copyright © 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd

This study was conducted to compare the effects of thermal stress on growth performance and some immunity variables of broiler chickens. Birds were randomly assigned to one of three thermal treatments as follows: cold stress (CS, 12±1°C), (b) heat stress (HS, 33±3 °C) and (c) thermoneutral (TN, 24±2 °C). Body weight gain (BWG), feed intake (FI), water intake (WI) and feed conversion ratio (FCR) were recorded. In order to evaluate the primary and secondary humoral immune responses, two birds per replicate were intravenously administrated with a suspension of 7% sheep red blood cell (SRBC) at 28 and 35 days. The heat-stressed broiler chickens had lower FI (-14.90%), BWG (-25.71%) and higher FCR (+13.06%) in comparison to broiler chickens reared under TN condition (p<0.001) from 1 to 42 days of age. The cold-stressed broiler chickens showed lower FI (-22.05%), BWG (-38.32%) and higher FCR (+22.47%) in comparison to birds reared under TN conditions (p<0.001). Stressed birds (CS and HS) showed decreased antibody titer against SRBC, lymphocyte count and the relative weights of lymphoid organs and increased heterophil count, heterophil to lymphocyte ratio and the serum concentration of corticosterone, in comparison to birds in TN group (p<0.001). In conclusion, HS and CS conditions have similar negative effects on performance and immunity of broiler chickens.

This study was conducted to compare the effects of thermal stress on growth performance and some immunity variables of broiler chickens. Birds were randomly assigned to one of three thermal treatments as follows: cold stress (CS, 12±1°C), (b) heat stress (HS, 33±3 °C) and (c) thermoneutral (TN, 24±2 °C). Body weight gain (BWG), feed intake (FI), water intake (WI) and feed conversion ratio (FCR) were recorded. In order to evaluate the primary and secondary humoral immune responses, two birds per replicate were intravenously administrated with a suspension of 7% sheep red blood cell (SRBC) at 28 and 35 days. The heat-stressed broiler chickens had lower FI (-14.90%), BWG (-25.71%) and higher FCR (+13.06%) in comparison to broiler chickens reared under TN condition (p<0.001) from 1 to 42 days of age. The cold-stressed broiler chickens showed lower FI (-22.05%), BWG (-38.32%) and higher FCR (+22.47%) in comparison to birds reared under TN conditions (p<0.001). Stressed birds (CS and HS) showed decreased antibody titer against SRBC, lymphocyte count and the relative weights of lymphoid organs and increased heterophil count, heterophil to lymphocyte ratio and the serum concentration of corticosterone, in comparison to birds in TN group (p<0.001). In conclusion, HS and CS conditions have similar negative effects on performance and immunity of broiler chickens.

Rheumatoid meningitis is a rare and very serious extra-articular manifestation of rheumatoid arthritis. We present a case of a 7()year-old female with no history of arthritis who developed stroke-like symptoms, seizures, psychosis and compulsive behavior. Serial brain magnetic resonance images (MRI) over four months demonstrated progressive interhemispheric meningeal thickening. She had mild lymphocytic pleocytosis on the cerebrospinal fluid analysis and serum anti-cyclic citrullinated peptide antibodies resulted positive in high titers. She underwent a brain biopsy showing necrotizing granulomas consistent with rheumatoid meningitis. Her symptoms resolved with treatment with glucocorticoids and cyclophosphamide. She has not been diagnosed with rheumatoid arthritis even after 1 year of follow up. Clinicians should be aware of the possibility of rheumatoid meningitis without rheumatoid arthritis and keep it on the differential for patients with aseptic meningitis and otherwise negative work up.

Thrombotic microangiopathy (TMA) is a well-recognised complication following transplantation, often due to an underlying genetic predisposition, medications or rejection. The use of eculizumab in these settings has been previously described, but its role still remains to be clarified. A 45-year-old man, with a history of type 1 diabetes mellitus and subsequent end-stage kidney failure, presented for a simultaneous pancreas-kidney transplant. Immunologically, he was well matched with the donor, and he received standard induction immunosuppression including tacrolimus. His early transplant course was complicated by Haemophilus parainfluenzae paronychia and a Pseudomonas aeruginosa catheter-associated urinary tract infection. Within 1 week, he developed thrombotic microangiopathy with significant renal dysfunction and eventual dialysis dependence, without evidence of transplant rejection on biopsy. He was also noted to have antiphospholipid antibodies in moderate titres. The TMA did not resolve despite cessation of tacrolimus, and he was subsequently commenced on eculizumab. The patient achieved a partial remission from TMA, with ongoing biochemical evidence of haemolysis, although now with stable graft function, despite significant damage. His transplanted pancreas remained seemingly unaffected by TMA, and continues to function well. This case describes an unusual presentation of TMA post-transplantation and is the only described case of eculizumab use following pancreas-kidney transplant. It remains unclear in this case what the likely precipitant for TMA was, although it seems to be, at least in part, controlled by ongoing use of eculizumab, presumably by terminal complement inhibition.Copyright © 2017 Asian Pacific Society of Nephrology

Thrombotic microangiopathy (TMA) is a well-recognised complication following transplantation, often due to an underlying genetic predisposition, medications or rejection. The use of eculizumab in these settings has been previously described, but its role still remains to be clarified. A 45-year-old man, with a history of type 1 diabetes mellitus and subsequent end-stage kidney failure, presented for a simultaneous pancreas-kidney transplant. Immunologically, he was well matched with the donor, and he received standard induction immunosuppression including tacrolimus. His early transplant course was complicated by Haemophilus parainfluenzae paronychia and a Pseudomonas aeruginosa catheter-associated urinary tract infection. Within 1 week, he developed thrombotic microangiopathy with significant renal dysfunction and eventual dialysis dependence, without evidence of transplant rejection on biopsy. He was also noted to have antiphospholipid antibodies in moderate titres. The TMA did not resolve despite cessation of tacrolimus, and he was subsequently commenced on eculizumab. The patient achieved a partial remission from TMA, with ongoing biochemical evidence of haemolysis, although now with stable graft function, despite significant damage. His transplanted pancreas remained seemingly unaffected by TMA, and continues to function well. This case describes an unusual presentation of TMA post-transplantation and is the only described case of eculizumab use following pancreas-kidney transplant. It remains unclear in this case what the likely precipitant for TMA was, although it seems to be, at least in part, controlled by ongoing use of eculizumab, presumably by terminal complement inhibition.Copyright © 2017 Asian Pacific Society of Nephrology

Aim Interactions between the co-stimulatory molecule programmed death 1 (PD-1) and its ligands, PD-L1 and PD-L2, constrain T-cell responses and help maintain peripheral tolerance. Glomerulonephritis can result from a variety of antigens, both self and foreign, and from humoural and cellular effector responses. These studies aimed to define the role of PD1 and its ligands in circulating immune complex glomerulonephritis induced by immunity to a foreign antigen. Methods Immune complex glomerulonephritis was initiated by injecting BALB/c mice with horse spleen apoferritin intraperitoneally daily for 14 days. Inhibitory anti-mouse PD-1, anti-PD-L1 or anti-PD-L2 antibodies were administered every other day. Renal disease and immune responses were studied. Results Daily injection of horse spleen apoferritin-induced proliferative immune complex glomerulonephritis in control antibody-treated mice, but inhibiting PD-1 did not augment renal injury. Specifically, blocking PD-1 did not increase serum antigen-specific antibodies or increase glomerular immunoglobulin G deposition, the hallmark of injury in this model. Furthermore, C3 deposition was unaffected and glomerular macrophages were reduced after anti-PD-1 antibodies. However, anti-PD-1 administration did increase splenocyte proliferation and cytokine production including interferon-gamma, interleukin (IL)-4, and IL-17, but not IL-10. Neutralizing either PD-L1 or PD-L2 alone did not result in major alterations in renal injury. Conclusion The endogenous PD-1/PD-L pathway does not limit acute experimental foreign antigen-induced circulating immune complex glomerulonephritis.Copyright © 2015 Asian Pacific Society of Nephrology.

Heparin-induced thrombocytopenia (HIT) and thrombosis is a serious adverse event associated with the widespread use of heparin as an inexpensive, fast-acting and reversible anticoagulant. The final diagnosis of HIT is still based on clinical suspicion as not all heparin/ PF4 antibodies cause HIT, and judgment from currently available assays for HIT-related Ig or its functional effects on platelets may be negative despite a convincing clinical picture. Consequently, it is difficult to distinguish HIT patients at risk of thrombosis from nonthrombotic HIT. Our previous studies showed that the plateletspecific receptor, glycoprotein (GP)VI, is stable on normal circulating platelets, but undergoes metalloproteinase-mediated ectodomain shedding in response to FclambdaRIIa-mediated platelet activation by HIT antibodies in vitro. We assessed whether soluble GPVI (sGPVI) was an early biomarker of HIT pathology by using an enzyme-linked immunosorbent assay (ELISA) to measure shed sGPVI in plasma from patients at risk or confirmed to have HIT. sGPVI levels in plasma from 192 healthy individuals [19.5+/-15.4 ng/mL] were independent of age and gender. sGPVI levels in HIT patients ranged from normal to > 180 ng/mL (mean 81+/-37.9 ng/mL) in plasma collected at the onset of thrombocytopenia. In 159 suspected HIT patients and 92 patients with thrombocytopenia unrelated to heparin, sGPVI levels were significantly higher in HIT-positive patients compared to thrombocytopenia or HIT-negative controls, with strong correlation with HIT-related antibody titre and capacity of patient plasma1heparin to induce aggregation of healthy donor platelets. These findings suggest a potential correlation between HIT-related Ig and plasma levels of sGPVI in vivo, consistent with previous in vitro experimental data and suggest that measurement of sGPVI may represent a useful adjunct assay for assessment of functional HIT.

Anti-neutrophil cytoplasmic antibody (ANCA)-associated glomerulonephritis involves innate and adaptive immune cells in the induction of autoimmunity and in autoimmune effector responses. Most Fcgamma receptors (FcgammaRs) activate immune cells, but FcgammaRIIB, found in humans and mice on B cells and innate cells, is an inhibitory receptor. Here we tested whether endogenous FcgammaRIIB negatively regulates autoreactivity and effector responses in experimental anti-myeloperoxidase (MPO) glomerulonephritis, using wild-type and FcgammaRIIB -/- mice. After MPO immunization, FcgammaRIIB -/- mice developed higher MPO-ANCA titers and increased anti-MPO T-cell responses. Transfer of FcgammaRIIB-deficient dendritic cells loaded with a nephritogenic MPO peptide (MPO 409-428) into wild-type mice induced stronger autoimmunity than dendritic cells derived from wild-type mice. Transferring anti-MPO antibodies into lipopolysaccharide-primed mice resulted in increased glomerular neutrophil accumulation and injury in FcgammaRIIB -/- mice, showing a role for FcgammaRIIB in suppressing neutrophil activation. Inducing active autoimmunity to MPO followed by triggering T cell-mediated glomerular injury by transfer of sub-nephritogenic doses of lipopolysaccharide and anti-MPO antibodies resulted in more disease in FcgammaRIIB -/- mice. Thus, endogenous FcgammaRIIB negatively regulates anti-MPO autoimmunity and glomerulonephritis by dendritic cells, B cells, and neutrophils to limit MPO-ANCA production, T-cell responses, and neutrophil activation.Copyright © 2014 International Society of Nephrology.

Aim Interactions between the co-stimulatory molecule programmed death 1 (PD-1) and its ligands, PD-L1 and PD-L2, constrain T-cell responses and help maintain peripheral tolerance. Glomerulonephritis can result from a variety of antigens, both self and foreign, and from humoural and cellular effector responses. These studies aimed to define the role of PD1 and its ligands in circulating immune complex glomerulonephritis induced by immunity to a foreign antigen. Methods Immune complex glomerulonephritis was initiated by injecting BALB/c mice with horse spleen apoferritin intraperitoneally daily for 14 days. Inhibitory anti-mouse PD-1, anti-PD-L1 or anti-PD-L2 antibodies were administered every other day. Renal disease and immune responses were studied. Results Daily injection of horse spleen apoferritin-induced proliferative immune complex glomerulonephritis in control antibody-treated mice, but inhibiting PD-1 did not augment renal injury. Specifically, blocking PD-1 did not increase serum antigen-specific antibodies or increase glomerular immunoglobulin G deposition, the hallmark of injury in this model. Furthermore, C3 deposition was unaffected and glomerular macrophages were reduced after anti-PD-1 antibodies. However, anti-PD-1 administration did increase splenocyte proliferation and cytokine production including interferon-gamma, interleukin (IL)-4, and IL-17, but not IL-10. Neutralizing either PD-L1 or PD-L2 alone did not result in major alterations in renal injury. Conclusion The endogenous PD-1/PD-L pathway does not limit acute experimental foreign antigen-induced circulating immune complex glomerulonephritis.Copyright © 2015 Asian Pacific Society of Nephrology.

Heparin-induced thrombocytopenia (HIT) and thrombosis is a serious adverse event associated with the widespread use of heparin as an inexpensive, fast-acting and reversible anticoagulant. The final diagnosis of HIT is still based on clinical suspicion as not all heparin/ PF4 antibodies cause HIT, and judgment from currently available assays for HIT-related Ig or its functional effects on platelets may be negative despite a convincing clinical picture. Consequently, it is difficult to distinguish HIT patients at risk of thrombosis from nonthrombotic HIT. Our previous studies showed that the plateletspecific receptor, glycoprotein (GP)VI, is stable on normal circulating platelets, but undergoes metalloproteinase-mediated ectodomain shedding in response to FclambdaRIIa-mediated platelet activation by HIT antibodies in vitro. We assessed whether soluble GPVI (sGPVI) was an early biomarker of HIT pathology by using an enzyme-linked immunosorbent assay (ELISA) to measure shed sGPVI in plasma from patients at risk or confirmed to have HIT. sGPVI levels in plasma from 192 healthy individuals [19.5+/-15.4 ng/mL] were independent of age and gender. sGPVI levels in HIT patients ranged from normal to > 180 ng/mL (mean 81+/-37.9 ng/mL) in plasma collected at the onset of thrombocytopenia. In 159 suspected HIT patients and 92 patients with thrombocytopenia unrelated to heparin, sGPVI levels were significantly higher in HIT-positive patients compared to thrombocytopenia or HIT-negative controls, with strong correlation with HIT-related antibody titre and capacity of patient plasma1heparin to induce aggregation of healthy donor platelets. These findings suggest a potential correlation between HIT-related Ig and plasma levels of sGPVI in vivo, consistent with previous in vitro experimental data and suggest that measurement of sGPVI may represent a useful adjunct assay for assessment of functional HIT.

Anti-neutrophil cytoplasmic antibody (ANCA)-associated glomerulonephritis involves innate and adaptive immune cells in the induction of autoimmunity and in autoimmune effector responses. Most Fcgamma receptors (FcgammaRs) activate immune cells, but FcgammaRIIB, found in humans and mice on B cells and innate cells, is an inhibitory receptor. Here we tested whether endogenous FcgammaRIIB negatively regulates autoreactivity and effector responses in experimental anti-myeloperoxidase (MPO) glomerulonephritis, using wild-type and FcgammaRIIB -/- mice. After MPO immunization, FcgammaRIIB -/- mice developed higher MPO-ANCA titers and increased anti-MPO T-cell responses. Transfer of FcgammaRIIB-deficient dendritic cells loaded with a nephritogenic MPO peptide (MPO 409-428) into wild-type mice induced stronger autoimmunity than dendritic cells derived from wild-type mice. Transferring anti-MPO antibodies into lipopolysaccharide-primed mice resulted in increased glomerular neutrophil accumulation and injury in FcgammaRIIB -/- mice, showing a role for FcgammaRIIB in suppressing neutrophil activation. Inducing active autoimmunity to MPO followed by triggering T cell-mediated glomerular injury by transfer of sub-nephritogenic doses of lipopolysaccharide and anti-MPO antibodies resulted in more disease in FcgammaRIIB -/- mice. Thus, endogenous FcgammaRIIB negatively regulates anti-MPO autoimmunity and glomerulonephritis by dendritic cells, B cells, and neutrophils to limit MPO-ANCA production, T-cell responses, and neutrophil activation.Copyright © 2014 International Society of Nephrology.