Search

BACKGROUND While B cell depletion is associated with attenuated antibody responses to SARS-CoV-2 mRNA vaccination, responses vary among individuals. Thus, elucidating the factors that affect immune responses after repeated vaccination is an important clinical need.METHODS We evaluated the quality and magnitude of the T cell, B cell, antibody, and cytokine responses to a third dose of BNT162b2 or mRNA-1273 mRNA vaccine in patients with B cell depletion.RESULTS In contrast with control individuals (n = 10), most patients on anti-CD20 therapy (n = 48) did not demonstrate an increase in spike-specific B cells or antibodies after a third dose of vaccine. A third vaccine elicited significantly increased frequencies of spike-specific non-naive T cells. A small subset of B cell–depleted individuals effectively produced spike-specific antibodies, and logistic regression models identified time since last anti-CD20 treatment and lower cumulative exposure to anti-CD20 mAbs as predictors of those having a serologic response. B cell–depleted patients who mounted an antibody response to 3 vaccine doses had persistent humoral immunity 6 months later.CONCLUSION These results demonstrate that serial vaccination strategies can be effective for a subset of B cell–depleted patients.FUNDING The NIH (R25 NS079193, P01 AI073748, U24 AI11867, R01 AI22220, UM 1HG009390, P01 AI039671, P50 CA121974, R01 CA227473, U01CA260507, 75N93019C00065, K24 AG042489), NIH HIPC Consortium (U19 AI089992), the National Multiple Sclerosis Society (CA 1061-A-18, RG-1802-30153), the Nancy Taylor Foundation for Chronic Diseases, Erase MS, the Robert Leet and Clara Guthrie Patterson Trust, and the Claude D. Pepper Older Americans Independence Center at Yale (P30 AG21342).

The skin lesion erythema migrans (EM) is an initial sign of the Ixodes tick–transmitted Borreliella spirochetal infection known as Lyme disease. T cells and innate immune cells have previously been shown to predominate the EM lesion and promote the reaction. Despite the established importance of B cells and antibodies in preventing infection, the role of B cells in the skin immune response to Borreliella is unknown. Here, we used single-cell RNA-Seq in conjunction with B cell receptor (BCR) sequencing to immunophenotype EM lesions and their associated B cells and BCR repertoires. We found that B cells were more abundant in EM in comparison with autologous uninvolved skin; many were clonally expanded and had circulating relatives. EM-associated B cells upregulated the expression of MHC class II genes and exhibited preferential IgM isotype usage. A subset also exhibited low levels of somatic hypermutation despite a gene expression profile consistent with memory B cells. Our study demonstrates that single-cell gene expression with paired BCR sequencing can be used to interrogate the sparse B cell populations in human skin and reveals that B cells in the skin infection site in early Lyme disease expressed a phenotype consistent with local antigen presentation and antibody production.

West Nile virus (WNV) infection is mainly asymptomatic but can be severe in elderly persons. As part of studies on immunity and aging in Connecticut, USA, we detected WNV seroconversion in 8.5% of nonimmunosuppressed and 16.8% of immunosuppressed persons. Age was not a significant seroconversion factor. Our findings suggest that immune factors affect seroconversion.

The West Nile virus (WNV) is an emerging infection of biodefense concern and there are no available treatments or vaccines. Here we used a high-throughput method based on a novel gene expression analysis, RNA-Seq, to give a global picture of differential gene expression by primary human macrophages of 10 healthy donors infected in vitro with WNV. From a total of 28 million reads per sample, we identified 1,514 transcripts that were differentially expressed after infection. Both predicted and novel gene changes were detected, as were gene isoforms, and while many of the genes were expressed by all donors, some were unique. Knock-down of genes not previously known to be associated with WNV resistance identified their critical role in control of viral infection. Our study distinguishes both common gene pathways as well as novel cellular responses. Such analyses will be valuable for translational studies of susceptible and resistant individuals—and for targeting therapeutics—in multiple biological settings.

Background: Around the world, individuals are living longer, but an increased average lifespan does not always equate to an increased healthspan. With advancing age, the increased prevalence of ageing-related diseases can have a significant impact on health status, functional capacity, and quality of life. It is therefore vital to develop comprehensive classification and staging systems for ageing-related pathologies, diseases and syndromes. This will allow societies to better identify, quantify, understand, and meet the healthcare, workforce, wellbeing, and socioeconomic needs of ageing populations, while supporting the development and utilisation of interventions to prevent or to slow, halt or reverse the progression of ageing-related pathologies., Methods: The foundation for developing such classification and staging systems is to define the scope of what constitutes an ageing-related pathology, disease or syndrome. To this end, a consensus meeting was hosted by the International Consortium to Classify Ageing-Related Pathologies (ICCARP), on February 19 th , 2024, in Cardiff, UK, and was attended by 150 recognised experts. Discussions and voting were centred on provisional criteria that had been distributed prior to the meeting. The participants debated and voted on these. Each criterion required a consensus agreement of ≥70% for approval., Results: The accepted criteria for an ageing-related pathology, disease or syndrome were: Develops and/or progresses with increasing chronological age.Should be associated with, or contribute to, functional decline, or an increased susceptibility to functional decline.Evidenced by studies in humans., Conclusions: Criteria for an ageing-related pathology, disease or syndrome have been agreed by an international consortium of subject experts. These criteria will now be used by the ICCARP for the classification and ultimately staging of ageing-related pathologies, diseases and syndromes.

High-dimensional mass cytometry data potentially enable a comprehensive characterization of immune cells. In order to positively affect clinical trials and translational clinical research, this advanced technology needs to demonstrate a high reproducibility of results across multiple sites for both peripheral blood mononuclear cells (PBMC) and whole blood preparations. A dry 30-marker broad immunophenotyping panel and customized automated analysis software were recently engineered and are commercially available as the Fluidigm® Maxpar® Direct™ Immune Profiling Assay™. In this study, seven sites received whole blood and six sites received PBMC samples from single donors over a 2-week interval. Each site labeled replicate samples and acquired data on Helios™ instruments using an assay-specific acquisition template. All acquired sample files were then automatically analyzed by Maxpar Pathsetter™ software. A cleanup step eliminated debris, dead cells, aggregates, and normalization beads. The second step automatically enumerated 37 immune cell populations and performed label intensity assessments on all 30 markers. The inter-site reproducibility of the 37 quantified cell populations had consistent population frequencies, with an average %CV of 14.4% for whole blood and 17.7% for PBMC. The dry reagent coupled with automated data analysis is not only convenient but also provides a high degree of reproducibility within and among multiple test sites resulting in a comprehensive yet practical solution for deep immune phenotyping., (© 2019 International Clinical Cytometry Society.)

It is currently challenging to analyze single-cell data consisting of many cells and samples, and to address variations arising from batch effects and different sample preparations. For this purpose, we present SAUCIE, a deep neural network that combines parallelization and scalability offered by neural networks, with the deep representation of data that can be learned by them to perform many single-cell data analysis tasks. Our regularizations (penalties) render features learned in hidden layers of the neural network interpretable. On large, multi-patient datasets, SAUCIE's various hidden layers contain denoised and batch-corrected data, a low-dimensional visualization and unsupervised clustering, as well as other information that can be used to explore the data. We analyze a 180-sample dataset consisting of 11 million T cells from dengue patients in India, measured with mass cytometry. SAUCIE can batch correct and identify cluster-based signatures of acute dengue infection and create a patient manifold, stratifying immune response to dengue.

Objective: The purpose of this article is to establish a body of literature exploring the emergent topic of Indigenous doulas, in relation to Indigenous communities in remote locations, where women are routinely evacuated and no longer supported to give birth. In doing so the article will synthesise and critique key concepts in the literature and identify gaps for prioritisation in future research., Design: The methodology is influenced by Indigenous, decolonising and feminist theoretical standpoints. A combined methodological approach of an integrative and scoping literature review was undertaken. Only published research, grey literature and grey data written in English and created between the years 2000 and 2018 was included. The search engines used were CINAHL plus, MEDLINE full text, Informat, Cochrane, Google Scholar and Google Search., Setting: Resources originating from only Canada and America identified and despite regional similarities, no literature from Australia or Greenland was sourced., Participants: Of the entirety of identified resources two author's Indigenous identity was readily identifiable; and in the research articles there was a total of 191 research participants identified as Indigenous. Much of the grey literature and grey data included quotations from Indigenous women., Interventions (if Appropriate): N/A., Measurements and Findings: Key concepts about the role and practice of Indigenous doulas were identified: reclaiming and supporting cultural practices; sovereignty over lands and bodies; strengthening families, training, work models and defiance of evacuation policies on the pathway to returning birth. Critique of these concepts suggests that Indigenous doulas have a unique role and practice scope in Western maternity care, which is readily distinguished from standard doula practice. Research gaps worthy of future research prioritisation include: Indigenous women's perspectives as recipients of Indigenous doula care, Indigenous doulas as a pathway into midwifery, escort policy and impacts on Indigenous doula provision; evaluation and alternative research settings., Key Conclusions and Implications for Practice: The role and practice of Indigenous doulas offers a promising approach to redressing the colonisation of Indigenous childbirth while contributing to improving Indigenous maternal and infant outcomes. Indigenous doula practice shares many best-practice characteristics with Indigenous Healing Programs and as such is also likely to also promote inter-generational healing. Most of the resources located were descriptive, but this emergent topic is worthy of further applied research., (Copyright © 2019. Published by Elsevier Ltd.)

The purpose of this study was to implement a sexual health behavioural intervention in Greenland in order to reduce sexually transmitted infection rates among a population of Greenland youth. This behavioural intervention was called Inuulluataarneq (Having the Good Life). Inuulluataarneq's objects included: (1) increase Greenlandic youth's overall knowledge about sexually transmitted infections and sexual health; (2) increase parent/guardian-youth communication about topics related to sexually transmitted infections and sex; and (3) increase consistent condom use among Greenlandic youth. We hypothesised that increased awareness of sexually transmitted infections and sexual health as well as increased communication between parents/guardians and their adolescent children would influence sexual risk behaviour and reduce sexually transmitted infections among our sample population, with a focus on urine samples of chlamydia infection. Results indicate that the influence of having a parent/guardian to speak with about topics related to sex, including the consequences of pregnancy, are key protective factors in reducing sexually transmitted infections among Greenlandic youth. Inuulluataarneq demonstrates that intensive short-term education and skill-building delivered by a trained community member is an effective sexually transmitted infection prevention intervention method among young Inuit populations who live in small isolated Arctic communities., (© The Author(s) 2014 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.)

Bacterial vaginosis (BV) is traditionally diagnosed using vaginal samples. The aim of this study was to investigate whether BV can be diagnosed from first-void urine (FVU). Self-collected vaginal smears, vaginal swabs, and FVU were obtained from 176 women. BV was diagnosed by Nugent's criteria. The FVU and vaginal swabs were analyzed by quantitative PCRs (qPCRs) for selected vaginal bacteria (Atopobium vaginae, Prevotella spp., Gardnerella vaginalis, bacterial vaginosis-associated bacterium 2, Eggerthella-like bacterium, "Leptotrichia amnionii," Megasphaera type 1), and all had an area under the receiver operating characteristic (ROC) curve of >85%, suggesting good prediction of BV according to the Nugent score. All seven bacteria in FVU were significantly associated with BV in univariate analysis. An accurate diagnosis of BV from urine was obtained in this population by a combination of qPCRs for Megasphaera type 1 and Prevotella spp. The same two bacteria remained significantly associated with BV in a multivariate model after adjusting for the other five species. There was no statistically significant difference between the sensitivities and specificities of BV diagnosis by molecular methods performed on swabs and FVU samples. A linear regression analysis showed good agreement between bacterial loads from swabs and FVU, but Prevotella spp. could be detected in high numbers in a few FVU samples without being present in swabs. This method will allow diagnosis of BV in studies where only urine has been collected and where detection of BV is considered relevant.

Induction of type I interferon is a central event of innate immunity, essential for host defense. Here we report that the transcription factor ELF4 is induced by type I interferon and upregulates interferon expression in a feed-forward loop. ELF4 deficiency leads to reduced interferon production, resulting in enhanced susceptibility to West Nile virus encephalitis in mice. After viral infection, ELF4 is recruited by STING, interacts with and is activated by the MAVS-TBK1 complex, and translocates into the nucleus to bind interferon promoters. Cooperative binding with ELF4 increases the binding affinity of interferon regulatory factors IRF3 and IRF7, which is mediated by EICE elements. Thus, in addition to identifying a regulator of innate immune signaling, we uncovered a role for EICE elements in interferon transactivation.

Objective: to review literature on the physical place of childbirth in Greenland between 1953 and 2001, using a narrative review theory and a content analysis framework, the paper seeks to describe and analyse the change in perinatal health care structure in Greenland., Design: findings were discussed within the framework of Daviss' Logics bringing into account scientific, clinical, personal, cultural and intuitive logics as well as economic, legal and political 'logics' concerning perinatal health care policies., Setting: the literature study concerns the place of birth in Greenland, a self-governing constituency of 57,000 people, the world's largest island and with a predominately Inuit population with its own language and culture. Inuit population with its own language and culture., Findings: the place of birth in Greenland has changed and focus has moved from birth as a personal and community act to birth within the private and political arena. New policies and guidelines for pregnancy and childbearing decisions are seldom negotiated with the women, families and their communities., Conclusions: policy changes have an influence on the social and cultural development of Greenland and it poses a challenge and a counter weight to the political and economic limitations that the government works within. Women and children are vulnerable groups and are directly affected by the changing perinatal health care and policy. It is important that when changing policy, the women and their families are part of the dialogue around change., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Background: Bacterial vaginosis (BV) is a common condition, although its aetiology remains unexplained. The aim of this study was to analyse the composition of vaginal microbiota in women from Greenland to provide a quantitative description and improve the understanding of BV., Methods: Self-collected vaginal smears and swabs were obtained from 177 women. The vaginal smears were graded for BV according to Nugent's criteria. The vaginal swab samples were analysed by 19 quantitative PCRs (qPCRs) for selected vaginal bacteria and by PCR for four sexually transmitted infections (STIs)., Results: STIs were common: Mycoplasma genitalium 12%, Chlamydia trachomatis 7%, Neisseria gonorrhoeae 1%, and Trichomonas vaginalis 0.5%. BV was found in 45% of women, but was not associated with individual STIs. Seven of the 19 vaginal bacteria (Atopobium vaginae, Prevotella spp., Gardnerella vaginalis, BVAB2, Eggerthella-like bacterium, Leptotrichia amnionii, and Megasphaera type 1) had areas under the receiver operating characteristic (ROC) curve > 85%, suggesting they are good predictors of BV according to Nugent. Prevotella spp. had the highest odds ratio for BV (OR 437; 95% CI 82-2779) in univariate analysis considering only specimens with a bacterial load above the threshold determined by ROC curve analysis as positive, as well as the highest adjusted odds ratio in multivariate logistic regression analysis (OR 4.4; 95% CI 1.4-13.5). BV could be subdivided into clusters dominated by a single or a few species together., Conclusions: BV by Nugent score was highly prevalent. Two of seven key species (Prevotella spp. and A. vaginae) remained significantly associated with BV in a multivariate model after adjusting for other bacterial species. G. vaginalis and Prevotella spp. defined the majority of BV clusters.

We present the ethical challenges and lessons learned over the course of a four-year community-based participatory research (CBPR) project conducted on sexually transmitted infections (STIs) in Greenland. Specifically discussed is Inuulluataarneq-the "Having the Good Life" study. Inuulluataarneq is an interdisciplinary international, collaborative CBPR study involving the University of Toronto in Canada, the Greenlandic Medical Research Council, the Centre for Primary Care in Nuuk, the University of Greenland, local health partners and communities in Greenland, the Statens Serum Institut in Denmark, and Montana State University in the United States. Inuulluataarneq is the first CBPR project implemented in Greenland. Ethical issues discussed are: (1) the complexity of working with multiple institutional review boards on an international health research project using a CBPR framework; (2) unexpected influences on health policy; and (3) the dynamic of balancing community decision making and practices with academic research requirements and expectations. Inuulluataarneq's primary contribution to understanding ethical issues when conducting research in the Arctic involves an acceptance of the time, patience, and dedication of researchers and community partners it takes to discuss, understand, and process differing ethical viewpoints and procedures.

Rationale: Lymphocytes are increasingly associated with idiopathic pulmonary fibrosis (IPF). Semaphorin 7a (Sema 7a) participates in lymphocyte activation., Objectives: To define the relationship between Sema 7a and lymphocytes in IPF., Methods: We characterized the significance of Sema 7a+ lymphocytes in humans with IPF and in a mouse model of lung fibrosis caused by lung-targeted, transgenic overexpression of TGF-β1. We determined the site of Sema 7a expression in human and murine lungs and circulation and used adoptive transfer approaches to define the relevance of lymphocytes coexpressing Sema7a and the markers CD19, CD4, or CD4+CD25+FoxP3+ in TGF-β1-induced murine lung fibrosis., Measurements and Main Results: Subjects with IPF show expression of Sema 7a on lung CD4+ cells and circulating CD4+ or CD19+ cells. Sema 7a expression is increased on CD4+ cells and CD4+CD25+FoxP3+ regulatory T cells, but not CD19+ cells, in subjects with progressive IPF. Sema 7a is expressed on lymphocytes expressing CD4 but not CD19 in the lungs and spleen of TGF-β1-transgenic mice. Sema 7a expressing bone marrow-derived cells induce lung fibrosis and alter the production of T-cell mediators, including IFN-γ, IL-4, IL-17A, and IL-10. These effects require CD4 but not CD19. In comparison to Sema 7a-CD4+CD25+FoxP3+ cells, Sema7a+CD4+CD25+FoxP3+ cells exhibit reduced expression of regulatory genes such as IL-10, and adoptive transfer of these cells induces fibrosis and remodeling in the TGF-β1-exposed murine lung., Conclusions: Sema 7a+CD4+CD25+FoxP3+ regulatory T cells are associated with disease progression in subjects with IPF and induce fibrosis in the TGF-β1-exposed murine lung.

Objectives: Greenland reports the highest rates of chlamydial infection and gonorrhea in the Arctic. Our objective was to determine the presence, and describe the basic epidemiology, of Mycoplasma genitalium for Greenland., Study Design: Cross-sectional study., Methods: 314 residents from Nuuk and Sisimiut, between the ages of 15 and 65 years, participated in "Inuulluataarneq" (the Greenland Sexual Health Project) between July 2008 and November 2009. Participants provided self-collected samples for sexually transmitted infection (STI) testing and completed a sexual health survey. Descriptive statistics and logistic regression were used to summarize the basic characteristics of STI cases overall and M. genitalium and Chlamydia trachomatis specifically. Clinically relevant characteristics in each full model were gender (male or female), age (in years), age at sexual debut (in years), number of sexual partners in the past 3 months (continuous) and history of forced sex and community., Results: The overall prevalence of STIs was 19.0%, specifically: 9.8% for M. genitalium and 9.4% for C. trachomatis; 100% of M. genitalium-positive cases carried macrolide resistance determinants. Being female [OR = 3.2; 95% confidence interval (CI): 1.1-9.8] and younger age (OR = 0.9; 95% CI: 0.9-1.0) were associated with M. genitalium positivity. Age was also associated with C. trachomatis (OR = 0.9; 95% CI: 0.8-0.9) and STI positivity overall (OR = 0.9; 95% CI: 0.9-0.9)., Conclusions: We observed a high prevalence of M. genitalium and macrolide resistance in this study. A better understanding of M. genitalium sequelae is needed to inform policy around testing, treatment, control and antibiotic use.

Objective: To study the effect of the innate cytokine macrophage migration inhibitory factor (MIF) on the susceptibility and severity of systemic lupus erythematosus (SLE) in a multinational population of 1,369 Caucasian and African American patients., Methods: Two functional polymorphisms in the MIF gene, a -794 CATT(5-8) microsatellite repeat (rs5844572) and a -173 G/C single-nucleotide polymorphism (rs755622), were assessed for association with SLE in 3,195 patients and healthy controls. We also measured MIF plasma levels in relation to genotypes and clinical phenotypes, and assessed Toll-like receptor 7 (TLR-7)-stimulated MIF production in vitro., Results: Both Caucasians and African Americans with the high-expression MIF haplotype -794 CATT(7)/-173*C had a lower incidence of SLE (in Caucasians, odds ratio [OR] 0.63, 95% confidence interval [95% CI] 0.53-0.89, P = 0.001; in African Americans, OR 0.46, 95% CI 0.23-0.95, P = 0.012). In contrast, among patients with established SLE, reduced frequencies of low-expression MIF genotypes (-794 CATT(5)) were observed in those with nephritis, those with serositis, and those with central nervous system (CNS) involvement when compared to patients without end-organ involvement (P = 0.023, P = 0.005, and P = 0.04, respectively). Plasma MIF levels and TLR-7-stimulated MIF production in vitro reflected the underlying MIF genotype of the studied groups., Conclusion: These findings suggest that MIF, which has both proinflammatory properties and macrophage and B cell survival functions, exerts a dual influence on the immunopathogenesis of SLE. High-expression MIF genotypes are associated with a reduced susceptibility to SLE and may contribute to an enhanced clearance of infectious pathogens. Once SLE develops, however, low-expression MIF genotypes may protect from ensuing inflammatory end-organ damage., (Copyright © 2011 by the American College of Rheumatology.)

Objectives: To develop a culturally competent and socially relevant sexual health survey for people living in Nuuk, Greenland, aged 15 years and older., Study Design: Qualitative study with interviews., Methods: Community and research informants (n=10) were interviewed informally to identify survey topics. A sexual health survey was constructed combining local knowledge from informants with a review of sexual health literature for the Arctic and other Indigenous locations. The draft survey was distributed to community partners for commentary and revision. After translation into Danish and Greenlandic, cognitive interviews were conducted with 11 Nuuk residents, identified through snowball sampling, to both pilot test the survey and exchange social and cultural knowledge relevant to sexual health in Nuuk. The utility of this process was evaluated against implementation of the final survey to Nuuk residents enrolled in Inuulluataarneq (n=149)., Results: Theme saturation was reached by the ninth interview. STI risk and self-efficacy, co-occurrence of alcohol use and sex and STI knowledge were identified as most relevant. Questions about community efficacy, culture/community involvement and identity were most sensitive. Upon implementation of the final survey, 146 of 149 participants answered all survey questions. Two Elder participants refused to answer questions about sex. Some questions had low response variability but still added to our contextual understanding and helped to build rapport with participants., Conclusions: Combining an iterative process with community-based participatory research principles and cognitive interview techniques was an effective method for developing a sexual health survey with Nuuk residents.

Increasingly, community-based participatory research (CBPR), with its emphasis on engaging communities as full and equal partners in all phases of the research process is being promoted to address the health needs of peoples living in the North American Arctic. However, the CBPR approach is not without its challenges in Arctic countries such as Greenland, where research capacity, different languages, distance, time and cost become barriers to remaining true to the purest form of CBPR. In this paper, we describe the practical application of CBPR principles and methodologies to a sexual health project investigating sexually transmitted infections in Greenland. We present the initial challenges encountered in the early stages of the pilot CBPR sexual health study, and solutions to these challenges. We also provide recommendations for expanding the capacity in Greenland to conduct CBPR projects.

Objectives: Adolescent pregnancy is a growing Public Health problem in Greenland, resulting in higher risk of mortality of mothers and their children. Since social and cultural aspects are associated with adolescent pregnancy, a closer look was taken at the situation of adolescent mothers in Greenland and in Native American communities., Methods and Results: Adolescent pregnancies and birth rates were followed in Greenland and in the First Nation communities in Alaska. Adolescent pregnancies decreased during the 1990s in both communities, but increased in 2000, bringing up the birth rate to 79 and 92 babies per 1,000 girls aged 15-19 yrs in Greenland in the U.S., respectively., Conclusions: A mentoring program to delay adolescent pregnancy and parenting, shown to be effective in African American and Latino communities, could be also used in the Greenlandic setting.

The GLH proteins belong to a family of four germline RNA helicases in Caenorhabditis elegans. These putative ATP-dependent enzymes localize to the P granules, which are nonmembranous complexes of protein and RNA exclusively found in the cytoplasm of all C. elegans germ cells and germ cell precursors. To determine what proteins the GLHs bind, C. elegans cDNA libraries were screened by the yeast two-hybrid method, using GLHs as bait. Three interacting proteins, CSN-5, KGB-1, and ZYX-1, were identified and further characterized. GST pull-down assays independently established that these proteins bind GLHs. CSN-5 is closely related to the subunit 5 protein of COP9 signalosomes, conserved multiprotein complexes of plants and animals. RNA interference (RNAi) with csn-5 results in sterile worms with small gonads and no oocytes, a defect essentially identical to that produced by RNAi with a combination of glh-1 and glh-4. KGB-1 is a putative JNK MAP kinase that GLHs bind. A kgb-1 deletion strain has a temperature-sensitive, sterile phenotype characterized by the absence of mature oocytes and the presence of trapped, immature oocytes that have undergone endoreplication. ZYX-1 is a LIM domain protein most like vertebrate Zyxin, a cytoskeletal adaptor protein. In C. elegans, while zyx-1 appears to be a single copy gene, neither RNAi depletion nor a zyx-1 deletion strain results in an obvious phenotype. These three conserved proteins are the first members in each of their families reported to associate with germline helicases. Similar to the loss of GLH-1 and GLH-4, loss of either CSN-5 or KGB-1 causes oogenesis to cease, but does not affect the initial assembly of P granules.