Biology subjects

Moriyama, M.

Publications and source records attributed to Moriyama, M..

17 recordsLinked to original sources

bioRxiv2026

Global transcriptional rewiring provides recurrent evolutionary routes to insect-Escherichia coli mutualism

Microbial symbionts often evolve specialized functions that compensate for host nutritional deficiencies, yet the evolutionary routes by which such mutualisms arise remain poorly understood. Here, using an experimental symbiosis between the stinkbug Plautia stali and Escherichia coli, we investigated the early evolution of insect-bacterium mutualism. Among 144 independently evolved bacterial lineages, six acquired the ability to improve host performance. Four lineages carried independent disruptive mutations in cyaA, disabling carbon catabolite repression and thereby increasing the essential amino acid tryptophan. The remaining two lineages evolved through nonsynonymous mutations in rpoB or rpoD, encoding core components of the transcriptional machinery. Transcriptomic analyses revealed extensive regulatory rewiring in both mutants, including marked upregulation of biosynthetic pathways for branched-chain essential amino acids. Our results identify global transcriptional regulatory systems as major evolutionary targets for the emergence of mutualism and demonstrate how large-effect regulatory mutations can rapidly generate host-beneficial traits. (146 < 150 words)

evolutionary biology↗

bioRxiv2026

Adaptive laboratory evolution of a minimal cell to low temperature

Temperature is a fundamental constraint on life. Most organisms adapt to temperature change through complex regulatory and stress-response systems. Whether such adaptation is possible in a genomically minimal cell remains unclear. Here, adaptive laboratory evolution was performed on the minimal genome bacterium JCVI-syn3B, shifting growth from 37{degrees}C to 25{degrees}C, a temperature at which the original strain cannot grow. After 40 serial passages, the evolved strains exhibited robust growth at 25{degrees}C. Multi-omics analyses revealed the adaptation mechanisms including increased mRNA turnover, enhanced DNA unwinding and stabilization of replication intermediates, elevated glycerolipid synthesis, and upregulation of division proteins associated with Z-ring assembly. Whole-genome transplantation was performed to distinguish genetic or non-genetic contributions, demonstrating that the cold-adapted phenotype was largely genetically encoded. These results indicated that even a minimal cell retains substantial evolvability and highlight its potential as a tractable research platform for cellular and evolutionary biology.

evolutionary biology↗

bioRxiv2026

Symbiotic Escherichia coli strains can better colonize host stinkbugs and outcompete natural symbiotic bacteria, but confer less fitness benefits

The stinkbug Plautia stali harbors essential gut symbiotic bacteria of the genus Pantoea, whose natural strains differ in cultivability and host benefits. Using this system, we evaluated how laboratory-evolved and genetically-engineered symbiotic Escherichia coli strains compete against native Pantoea symbionts and how they influence host fitness. In single infection assays, the native uncultivable symbiont Sym A conferred the highest host performance, whereas the evolved (CmL05G13) and artificial ({Delta}cyaA) symbiotic E. coli strains supported host survival at levels comparable to cultivable Pantoea symbionts (Sym C-F). In competitive co-infection assays, the symbiotic E. coli strains generally showed unexpectedly strong colonization ability. CmL05G13 outcompeted all the cultivable symbionts Sym C-F and even displaced the native uncultivable symbiont Sym A, whereas {Delta}cyaA and the nonsymbiotic control E. coli {Delta}intS were dominated by Sym A at the adult stage. Despite their superior infection competitiveness, the symbiotic E. coli strains provided limited reproductive benefits, behaving as "cheater-like" associates. They were able to invade and dominate the symbiotic organ but failed to match the fitness contributions of native symbionts. These results demonstrate that the experimentally evolved E. coli can rapidly acquire strong colonization ability surpassing that of the natural symbionts that have coevolved with P. stali in nature. At the same time, the mismatch between infection success and host fitness benefits highlights potential evolutionary conflicts and provides an experimental model for studying the dynamics of cheating, mutualism, and symbiont replacement in vertically transmitted symbioses. IMPORTANCEUnderstanding how novel symbionts invade and displace long-term mutualists is central to the evolution of symbiosis. This study demonstrates that Escherichia coli, originally a nonsymbiotic bacterium, can rapidly evolve potent colonization ability and even outcompete native Pantoea symbionts of the stinkbug Plautia stali. Meanwhile, these competitive E. coli strains confer markedly lower reproductive benefits compared with the native symbionts that have developed intimate mutualistic association with host P. stali over evolutionary time, revealing a striking decoupling between infection success and host fitness. This finding highlights the potential for cheater-like microbes to invade vertically transmitted symbioses and destabilize coevolved partnerships. By combining experimental evolution, controlled co-infections, and quantitative analyses, the P. stali-E. coli experimental symbiotic system provides a powerful model for studying the mechanisms and evolutionary dynamics of mutualism, cheating, and symbiont replacement.

microbiology↗

bioRxiv2025

Uric acid accumulation in cockroach wings

Cockroaches, embracing some 4,500 species in the world, are found in a variety of environments including human settlements. The ecological success of cockroaches is attributable to the ability to endure long periods of starvation and to thrive on nutritionally poor diets, which is underpinned by the capability of storing their own nitrogenous waste products as uric acid in the fat body and also by the endosymbiosis with bacteriocyte-dwelling bacteria Blattabacterium that recycle the stored uric acid for synthesis of amino acids. Previous histological studies on cockroaches described that uric acid is accumulated within the fat body, especially in urocytes. Here we report that cockroaches also accumulate uric acid in the wings. In our observation of the German cockroach Blattella germanica under different nutritional conditions, we found that well-fed cockroaches exhibited whitish aggregates in the wing veins, which is ascribed to accumulation of uric acid granules. The uric acid granules in the wings disappeared by starvation and were restored by feeding in B. germanica. The uric acid granules in the wings were found not only in B. germanica but also in other diverse cockroach species.

physiology↗

bioRxiv2025

Cockroach bacteriocytes migrate into the ovaries for vertical transmission of the bacterial endosymbiont Blattabacterium

Diverse insect groups are obligatorily associated with and dependent on specific microorganisms as essential mutualistic partners that are usually maintained in specialized cells or organs, called bacteriocytes or symbiotic organs. Many organisms with symbiotic microorganisms have developed elaborate vertical transmission mechanisms, which are thought to be important for the evolution of intimate symbiotic relationships with microorganisms. One such case is the cockroach-Blattabacterium endosymbiosis, in which the symbiotic bacteria have been evolutionarily conserved and co-speciated with the host insects with stable vertical symbiont transmission via ovarial passage. While classical histological descriptions and recent electron microscopic observations have reported the vertical symbiont transmission processes in some cockroach-Blattabacterium associations, the full picture of the infection dynamics has not been fully understood. In this study, we conducted detailed histological and cytological observations of the localization of the bacteriocytes and the symbiotic bacteria during the postembryonic development of the German cockroach Blattella germanica. We found that the symbiont-filled bacteriocytes migrate into and associate with nymphal ovaries and are subsequently eliminated from adult ovaries, suggesting that symbiont infection to the ovaries may only occur during nymphal stages. We also found that the symbiotic bacteria are localized in the space between each oocyte and surrounding follicle cells, the symbiont-localized space is interconnected between neighboring oocytes, and therefore the symbiotic bacteria can move across oocytes within the same ovariole, suggesting the possibility that the symbiont-infected oocytes may serve as the source of symbiont supply to developing young oocytes upstream in the same ovariole. Based on these observations, we provide a hypothesis as to how the postembryonic developmental dynamics of the bacteriocytes are integrated into the vertical symbiont transmission and functioning in the cockroach-Blattabacterium endosymbiosis.

developmental biology↗

bioRxiv2025

Acquisition and repeated alteration of (TTGGG)n telomeric repeats in Odonata (dragonflies and damselflies)

Chromosome ends of most eukaryotes are composed of simple telomeric repeats. For arthropods, TTAGG pentanucleotide repeats, (TTAGG)n has been considered as the ancestral telomeric repeat. However, in the order Odonata, the earliest diverged group in insects that contains dragonflies and damselflies, (TTAGG)n signal has been almost undetectable in most examined species. Here, we report a unique pentanucleotide repeat (TTGGG)n as the typical telomeric repeat sequence for Odonata (dragonflies and damselflies). Based on genomic information from ten Odonata species, (TTGGG)n was considered the most likely candidate for telomeric repeat sequences. By fluorescence in situ hybridization (FISH) using 12 Odonata species, clear (TTGGG)n signals were detected at the chromosome ends in both dragonflies and damselflies. By Southern hybridization using 63 Odonata species, strong (TTGGG)n signals were detected from the majority of species covering all three suborders of Odonata, indicating that the telomeric repeat of the common ancestor of extant Odonata is (TTGGG)n. Notably, there were a few Odonata species in which (TTGGG)n signals were faint or absent, suggesting that the telomeric repeat sequence has repeatedly diverged in Odonata, even within genera such as Sympetrum. We also identified telomerase genes in both dragonflies and damselflies, and in some species, more than two telomerase genes are suggested to be present. Overall, this study demonstrates the ancestral acquisition of novel telomeric repeats and their repeated alteration in Odonata.

cell biology↗

bioRxiv2024

Intranasal hemagglutinin protein boosters induce robust mucosal immunity and cross-protection against influenza A viral challenge

Licensed parenteral influenza vaccines induce systemic antibody responses and alleviate disease severity but do not efficiently prevent viral entry and transmission due to the lack of local mucosal immune responses. Here, we describe intranasal booster strategy with unadjuvanted recombinant hemagglutinin (HA) following initial mRNA-LNP vaccination, Prime and HA. This regimen establishes highly protective HA-specific mucosal immune memory responses in the respiratory tract. Intranasal HA boosters provided significantly reduced viral replication compared to parenteral mRNA-LNP boosters in both young and old mice. Correlation analysis revealed that slightly increased levels of nasal IgA are significantly associated with a reduced viral burden in the upper respiratory tract. Intranasal boosting with an antigenically distinct H1 HA conferred sterilizing immunity against heterologous H1N1 virus challenge. Additionally, a heterosubtypic intranasal H5 HA booster elicited cross-reactive mucosal humoral responses. Our work illustrates the potential of a nasal HA protein booster as a needle- and adjuvant-free strategy to prevent infection and disease from influenza A viruses. One Sentence SummaryAdjuvant-free nasal booster induces protective immunity against influenza infection.

immunology↗

bioRxiv2024

Tryptophanase disruption underlies the evolution of insect-bacterium mutualism

Animal-microbe symbioses are omnipresent, where both partners often gain benefits as mutualists. How such mutualism has evolved between originally unrelated organisms is of interest. Here we report that, using an experimental symbiotic system between the stinkbug Plautia stali and the model bacterium Escherichia coli, disruption of a single bacterial gene tnaA encoding tryptophanase makes E. coli mutualistic to P. stali. Survey of natural bacterial mutualists across wild populations of P. stali and other stinkbug species uncovered that their Pantoea-allied symbionts consistently lack tnaA gene. Some Pantoea species like P. ananatis retain tnaA gene and cannot establish symbiosis with P. stali, but tnaA-disrupted P. ananatis partially restored the symbiotic capability. When a natural Pantoea mutualist of P. stali was transformed with a functional tna operon, its symbiotic capability reduced significantly. Our finding suggests that tryptophanase disruption may have facilitated the evolution of gut bacterial mutualists in insects.

evolutionary biology↗

bioRxiv2024

Host range of naturally and artificially evolved symbiotic bacteria for a specific host insect

Diverse insects are intimately associated with specific symbiotic bacteria, where host and symbiont are integrated into an almost inseparable biological entity. These symbiotic bacteria usually exhibit host specificity, uncultivability, reduced genome size, and other peculiar traits relevant to their symbiotic lifestyle. How host-symbiont specificity is established at the very beginning of symbiosis is of interest but poorly understood. To gain insight into the evolutionary issue, we adopted an experimental approach using the recently developed evolutionary model of symbiosis between the stinkbug Plautia stali and Escherichia coli. Based on the laboratory evolution of P. stali-E. coli mutualism, we selected {Delta}cyaA mutant of E. coli as an artificial symbiont of P. stali that has established mutualism by a single mutation. In addition, we selected a natural cultivable symbiont of P. stali of relatively recent evolutionary origin. These artificial and natural symbiotic bacteria of P. stali were experimentally inoculated to symbiont-deprived newborn nymphs of diverse stinkbug species. Strikingly, the mutualistic E. coli was unable to establish infection and support growth and survival of all the stinkbug species except for P. stali, uncovering that host specificity can be established at a very early stage of symbiotic evolution. Meanwhile, the natural symbiont was able to establish infection and support growth and survival of several stinkbug species in addition to P. stali, unveiling that a broader host range of the symbiont has evolved in nature. Based on these findings, we discuss what factors are relevant to the establishment of host specificity in the evolution of symbiosis. IMPORTANCEHow does host-symbiont specificity emerge at the very beginning of symbiosis? This question is difficult to address, because it is generally difficult to directly observe the onset of symbiosis. However, recent development of experimental evolutionary approaches to symbiosis has brought about a breakthrough. Here we tackled this evolutionary issue using a symbiotic Escherichia coli created in laboratory and a natural Pantoea symbiont, which are both mutualistic to the stinkbug Plautia stali. We experimentally replaced essential symbiotic bacteria of diverse stinkbugs by the artificial and natural symbionts of P. stali, and evaluated whether the symbiotic bacteria evolved for a specific host can establish infection and support growth and survival of heterospecific hosts. Strikingly, the artificial symbiont showed strict host specificity to P. stali whereas the natural symbiont was capable of symbiosis with diverse stinkbugs, which provide insight into how host-symbiont specificity can be established at early evolutionary stages of symbiosis.

microbiology↗

bioRxiv2024

High temperature treatment optimized for symbiont suppression in an obligatory gut bacterial symbiosis in the stinkbug Plautia stali

In the era of global warming, much attention has been paid to the possibility that high temperature may influence diverse insects not only directly but also indirectly via effects on their symbiotic microorganisms. The stinkbug Plautia stali develops a midgut symbiotic organ that harbors a specific bacterial symbiont indispensable for its growth and survival. Being maintainable in laboratory and tractable experimentally, P. stali is recently highlighted as a model system to investigate the mechanisms underpinning insect-microbe symbiotic interactions. In this study, we reared newly-emerged adult insects of P. stali under different temperature conditions for 8 days and monitored how their symbiotic organs and symbiotic bacteria are affected. While all insects survived at temperatures from 25{degrees}C to 37{degrees}C, some insects died at 38{degrees}C, 39{degrees}C and 40{degrees}C, wherein mortality rates increased as temperature elevated. While the symbiotic organs of the normal insects exhibited vivid yellow color, the symbiotic organs of the insects reared at 35{degrees}C or higher frequently exhibited abnormal colors such as pale yellow, yellowish white, or white, the extent of which became more severe as temperature elevated. Symbiont quantification revealed that, while the symbiont titers were almost constant for 8 days at 25{degrees}C and 30{degrees}C, the symbiont titers on the 8th day drastically declined to 1/100 at 35 and 1/10000 at 37{degrees}C and 39{degrees}C. Based on these results, we propose that rearing at 37{degrees}C for a week is a recommended treatment regime by which the symbiont is effectively suppressed with minimal damage to the host insect.

physiology↗

bioRxiv2024

Defensive fungal symbiosis on insect hindlegs

Tympanal organs as "insect ears" have evolved repeatedly. Dinidorid stinkbugs were reported to possess a conspicuous tympanal organ on females hindlegs. Here we report an unexpected discovery that the stinkbugs "tympanal organ" is actually a novel symbiotic organ. The stinkbugs "tympanum" is not membranous but a porous cuticle, where each pore connects to glandular secretory cells. In reproductive females, the hindleg organ is covered with fungal hyphae growing out of the pores. Upon oviposition, the females skillfully transfer the fungi from the organ to the eggs. The eggs are quickly covered with hyphae and physically protected against wasp parasitism. The fungi are mostly benign Cordycipitaceae entomopathogens and show considerable diversity among insect individuals and populations, indicating environmental acquisition of specific fungal associates. These results uncover a novel external fungal symbiosis in which hosts elaborate morphological, physiological and behavioral specializations underpin the selective recruitment of benign entomopathogens for a defensive purpose.

microbiology↗

bioRxiv2023

A novel monocarboxylate transporter involved in 3-hydroxykynurenine transport for ommochrome coloration

Ommochromes are widespread pigments in invertebrates utilized for screening pigments in compound eyes and for reddish coloration in epidermis and wings. Ommochromes are derived from 3-hydroxykynurenine (3OHK), which is incorporated into cells from hemolymph or synthesized from tryptophan within cells. While the synthetic pathway from tryptophan to 3OHK has been well characterized, the gene responsible for cellular uptake of 3OHK has been poorly understood. In the silkworm Bombyx mori, adult compound eyes and eggs contain a mixture of ommochrome pigments. By using positional cloning method, we found that a novel monocarboxylate transporter, 3-hydroxykynurenine transporter (3OHKT), is responsible for the recessive mutant maternal brown of Tsujita (b-t) of B. mori. In b-t mutant, the color of the eggs is light brown, whereas the color of the compound eyes is normal, and we identified a 2-kb deletion in 3OHKT gene. TALEN-mediated knockout of 3OHKT gene produced the same coloration phenotype as b-t mutant, and the complementation test between b-t mutant and 3OHKT knockout strain proved that 3OHKT is responsible for b-t phenotype. 3OHKT protein was localized in the cellular membrane, and LC-MS analysis indicated that the uptake of 3OHK from hemolymph into the ovary was suppressed in the b-t mutant. Moreover, we confirmed that 3OHKT gene is specifically expressed at the reddish region and the time of pigmentation in the pupal wing of nymphalid butterflies. RNA interference of 3OHKT prevented reddish pigmentation in wings, highlighting its general involvement in ommochrome-based pigmentation other than compound eyes. SignificanceOmmochromes are widely distributed pigments in invertebrates and are synthesized from intracellular tryptophan or 3-hydroxykynurenine (3OHK). Ommochrome-based red markings on butterfly wings are often used for sexual selection, warning colors and mimicry. Most genes involved in the ommochrome synthesis pathway have been elucidated from analyses of eye color mutants in Drosophila. However, this study reveals that the ommochrome synthesis pathway has a different genetic repertoire depending on the tissues, and that the novel monocarboxylate transporter identified in this study has a major role in ommochrome pigmentation other than in compound eyes. In particular, our results suggest that classical ommochrome-related genes are rarely involved in the wing pigmentation of the nymphalid butterflies.

genetics↗

bioRxiv2023

Mechanisms underpinning morphogenesis of symbiotic organ specialized for hosting indispensable microbial symbiont in stinkbug

Microbial mutualists are pivotal for insect adaptation, which often entails the evolution of elaborate organs for symbiosis. Addressing what mechanisms underpin the development of such organs is of evolutionary interest. Here we investigated the stinkbug Plautia stali whose posterior midgut is transformed into a specialized symbiotic organ. Despite being a simple tube in newborns, it developed numerous crypts in four rows, whose inner cavity hosts a specific bacterial symbiont, during 1st to 2nd nymphal instar. Visualization of dividing cells revealed that active cell proliferation was coincident with the crypt formation, although spatial patterns of the proliferating cells did not reflect the crypt arrangement. Visualization of visceral muscles in the midgut, consisting of circular muscles and longitudinal muscles, uncovered that, strikingly, circular muscles exhibited a characteristic arrangement running between the crypts specifically in the symbiotic organ. Even in early 1st instar when no crypts were seen, two rows of epithelial areas delineated by bifurcated circular muscles were identified. In 2nd instar, crossing muscle fibers newly appeared and connected the adjacent circular muscles, whereby the midgut epithelium was divided into four rows of crypt-to-be areas. The crypt formation proceeded even in aposymbiotic nymphs, uncovering autonomous nature of the crypt development. We propose a mechanistic model of crypt formation wherein the spatial arrangement of muscle fibers and the proliferation of epithelial cells underpin the formation of crypts as midgut evaginations. IMPORTANCEDiverse organisms are associated with microbial mutualists, in which specialized host organs often develop for retaining the microbial partners. In the light of the origin of evolutionary novelties, it is important to understand what mechanisms underpin the elaborate morphogenesis of such symbiotic organs, which must have been shaped through interactions with the microbial symbionts. Using the stinkbug Plautia stali as a model, we demonstrated that visceral muscular patterning and proliferation of intestinal epithelial cells during early nymphal stages are involved in the formation of numerous symbiont-harboring crypts arranged in four rows in the posterior midgut to constitute the symbiotic organ. Strikingly, the crypt formation occurred normally even in symbiont-free nymphs, uncovering that the crypt development proceeds autonomously. These findings suggest that the crypt formation is deeply implemented into the normal development of P. stali, which must reflect the considerably ancient evolutionary origin of the midgut symbiotic organ in stinkbugs.

microbiology↗

bioRxiv2022

SARS-CoV-2 variants do not evolve to promote further escape from MHC-I recognition

SARS-CoV-2 variants of concern (VOCs) possess mutations that confer resistance to neutralizing antibodies within the Spike protein and are associated with breakthrough infection and reinfection. By contrast, less is known about the escape from CD8+ T cell-mediated immunity by VOC. Here, we demonstrated that all SARS-CoV-2 VOCs possess the ability to suppress MHC I expression. We identified several viral genes that contribute to the suppression of MHC I expression. Notably, MHC-I upregulation was strongly inhibited after SARS-CoV-2 infection in vivo. While earlier VOCs possess similar capacity as the ancestral strain to suppress MHC I, Omicron subvariants exhibit a greater ability to suppress surface MHC-I expressions. Collectively, our data suggest that, in addition to escape from neutralizing antibodies, the success of Omicron subvariants to cause breakthrough infection and reinfection may in part be due to its optimized evasion from T cell recognition. SignificanceNumerous pathogenic viruses have developed strategies to evade host CD8+ T cell-mediated clearance. Here, we demonstrated that SARS-CoV-2 encodes multiple viral factors that can modulate MHC-I expression in the host cells. We found that MHC-I upregulation was strongly suppressed during SARS-CoV-2 infection in vivo. Notably, the Omicron subvariants showed an enhanced ability to suppress MHC-I compared to the original strain and the earlier SARS-CoV-2 variants of concern (VOCs). Our results point to the inherently strong ability of SARS-CoV-2 to hinder MHC-I expression and demonstrated that Omicron subvariants have evolved an even more optimized capacity to evade CD8 T cell recognition.

microbiology↗

bioRxiv2022

Single mutation makes Escherichia coli an insect mutualist

We report an experimental system in which Escherichia coli evolves into an insect mutualist. When the essential gut symbiont of the stinkbug Plautia stali was replaced by E. coli, a few survivor insects exhibited specific localization and vertical transmission of E. coli. Through trans-generational maintenance with P. stali, several hyper-mutating E. coli lines independently evolved hosts high adult emergence and improved body color. Such "mutualistic" E. coli lines exhibited independent mutations disrupting the carbon catabolite repression (CCR) global transcriptional regulator. Each of the mutations reproduced the mutualistic phenotypes when introduced into wild-type E. coli, confirming that the single CCR mutations instantly make E. coli an insect mutualist. Our discovery uncovers that evolution of elaborate mutualism can proceed more easily and rapidly than conventionally envisaged.

evolutionary biology↗

bioRxiv2020

Disruption of nasal bacteria enhances protective immune responses to influenza A virus and SARS-CoV-2 infection in mice

Gut microbiota plays a critical role in the induction of adaptive immune responses to influenza virus infection. However, the role of nasal bacteria in the induction of the virus-specific adaptive immunity is less clear. Here we demonstrate that while intranasal administration of influenza virus hemagglutinin vaccine alone was insufficient to induce the vaccine-specific antibody responses, disruption of nasal bacteria by lysozyme or addition of culturable oral bacteria from a healthy human volunteer rescued inability of the nasal bacteria to generate antibody responses to intranasally administered the split-virus vaccine. Myd88-depdnent signaling in the hematopoietic compartment was required for adjuvant activity of intranasally administered oral bacteria. In addition, we found that the oral bacteria-combined intranasal vaccine induced protective antibody response to influenza virus and SARS-CoV-2 infection. Our findings here have identified a previously unappreciated role for nasal bacteria in the induction of the virus-specific adaptive immune responses.

microbiology↗

bioRxiv2018

The expansion in lymphoid organs of IL-4+ BATF+ T follicular helper cells is linked to IgG4 class switching in vivo

Distinct TFH subsets that influence specific class-switching events are assumed to exist, but the accumulation of isotype-specific TFH subsets in secondary and tertiary lymphoid organs has not been hitherto demonstrated. IL-4 expressing TFH cells are surprisingly sparse in human secondary lymphoid organs. In sharp contrast, in IgG4-related disease (IgG4-RD), a disorder characterized by polarized Ig class switching, most TFH cells in tertiary and secondary lymphoid organs make IL-4. Human IL-4+ TFH cells do not express GATA-3 but express nuclear BATF, and the transcriptomes of IL-4 secreting TFH cells differ both from PD1hi TFH cells that do not secrete IL-4 and IL4-secreting non-TFH cells. Unlike IgG4-RD, IL-4+ TFH cells are rarely found in tertiary lymphoid organs in Sjogrens syndrome, a disorder in which IgG4 is not elevated. The proportion of CD4+IL-4+BATF+ T cells as well as of CD4+IL-4+CXCR5+ T cells in IgG4-RD tissues correlates tightly with tissue IgG4 plasma cell numbers and plasma IgG4 levels in patients but not with the total plasma levels of other isotypes. These data describe a disease-related TFH sub-population in human tertiary and secondary lymphoid organs that is linked to IgG4 class switching.

immunology↗