Rapid and precise identification of immature stages of leafminers of the genus Liriomyza Mik associated with imported fresh produce is essential to ensure appropriate biosecurity decisions at the border, in quarantine and post border. The leafminers Liriomyza huidobrensis, Liriomyza sativae and Liriomyza trifolii are not present in New Zealand and classified as regulated pests when detected at New Zealand's border. To assist rapid species identification of the immature stages of interceptions, a multiplex real-time TaqMan PCR assay was developed to identify these three species simultaneously in a single test. Species-specific primers and probes were designed by amplifying the mitochondrial COI gene of each targeting species, respectively. The multiplex real-time PCR assay demonstrated high specificity for all three target species and the assay detected DNA quantities as low as 0.1 pg for all species. Linear responses and high correlation coefficients between the amount of DNA and C q values for each species were also achieved. Therefore, the assay demonstrated its sensitivity and reliability for the identification of these three invasive Liriomyza species.

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IntroductionInclusion body myositis (IBM) is a progressive inflammatory myopathy characterised by skeletal muscle infiltration and myofibre invasion by CD8+ T lymphocytes. In some cases, IBM has been reported to be associated with a systemic lymphoproliferative disorder of CD8+ T cells exhibiting a highly differentiated effector phenotype known as T cell Large Granular Lymphocytic Leukemia (T-LGLL). MethodsWe investigated the incidence of a CD8+ T-LGL lymphoproliferative disorder in 85 IBM patients and an aged-matched group of 56 Healthy Controls (HC). Further, we analysed the phenotypical characteristics of the expanded T-LGLs and investigated whether their occurrence was associated with any particular HLA alleles or clinical characteristics. ResultsBlood cell analysis by flow cytometry revealed expansion of T-LGLs in 34 of the 85 (40%) IBM patients. The T cell immunophenotype of T-LGLHIGH patients was characterised by increased expression of surface molecules including CD57 and KLRG1, and to a lesser extent of CD94 and CD56 predominantly in CD8+ T cells, although we also observed modest changes in CD4+ T cells and γδ T cells. Analysis of Ki67 in CD57+ KLRG1+ T cells revealed that only a small proportion of these cells was proliferating. Comparative analysis of CD8+ and CD4+ T cells isolated from matched blood and muscle samples donated by three patients indicated a consistent pattern of more pronounced alterations in muscles, although not significant due to small sample size. In the T-LGLHIGH patient group, we found increased frequencies of perforin-producing CD8+ and CD4+ T cells that were moderately correlated to combined CD57 and KLRG1 expression. Investigation of the HLA haplotypes of 75 IBM patients identified that carriage of the HLA-C*14:02:01 allele was significantly higher in T-LGLHIGH compared to T-LGLLOW individuals. Expansion of T-LGL was not significantly associated with seropositivity patient status for anti-cytosolic 5'-nucleotidase 1A autoantibodies. Clinically, the age at disease onset and disease duration were similar in the T-LGLHIGH and T-LGLLOW patient groups. However, metadata analysis of functional alterations indicated that patients with expanded T-LGL more frequently relied on mobility aids than T-LGLLOW patients indicating greater disease severity. ConclusionAltogether, these results suggest that T-LGL expansion occurring in IBM patients is correlated with exacerbated immune dysregulation and increased disease burden.

Pre-existing pathogen-specific memory T cell responses can contribute to multiple adverse outcomes including autoimmunity and drug hypersensitivity. How the specificity of the T cell receptor (TCR) is subverted or seconded in many of these diseases remains unclear. Here, we apply abacavir hypersensitivity (AHS) as a model to address this question because the disease is linked to memory T cell responses and the HLA risk allele, HLA-B*57:01, and the initiating insult, abacavir, are known. To investigate the role of pathogen-specific TCR specificity in mediating AHS we performed a genome-wide screen for HLA-B*57:01 restricted T cell responses to Epstein-Barr virus (EBV), one of the most prevalent human pathogens. T cell epitope mapping revealed HLA-B*57:01 restricted responses to 17 EBV open reading frames and identified an epitope encoded by EBNA3C. Using these data, we cloned the dominant TCR for EBNA3C and a previously defined epitope within EBNA3B. TCR specificity to each epitope was confirmed, however, cloned TCRs did not cross-react with abacavir plus self-peptide. Nevertheless, abacavir inhibited TCR interactions with their cognate ligands, demonstrating that TCR specificity may be subverted by a drug molecule. These results provide an experimental road map for future studies addressing the heterologous immune responses of TCRs including T cell mediated adverse drug reactions.

The New Zealand endemic Pachycladon exile is a weakly perennial analogue of the intensively studied Arabidopsis thaliana. Imbibed seeds of P. exile that had been subjected to different durations of chilling at 5 8C (30, 20, 10 and 0 days) were germinated and grown for 10 weeks under controlled conditions of photoperiod (long or short days) in order to determine the effect of environment on vegetative phase change and flowering. In addition, meristem identity gene equivalents of A. thaliana LEAFY and TERMINAL FLOWER1 were isolated from P. exile and characterized as a first step towards gene expression studies in this perennial species. Germination in P. exile was enhanced by moderate chilling (10Á20 days), and flowering was facultative to both long days and chilling. As in A. thaliana, the appearance of abaxial trichomes might serve as a phase change marker in P. exile.