Matthew Woodruff

Emory University | EU · Division of Rheumatology

Following infection or vaccination, early-minted antibody secreting cells (ASC) or plasmablasts appear in circulation transiently, and a small fraction migrates to the spleen or bone marrow (BM) to mature into long-lived plasma cells (LLPC). While LLPC, by definition, are quiescent or non-dividing, the majority of blood ASC are thought to be 'blast...

While immunologic correlates of COVID-19 have been widely reported, their associations with post-acute sequelae of COVID-19 (PASC) remain less clear. Due to the wide array of PASC presentations, understanding if specific disease features associate with discrete immune processes and therapeutic opportunities is important. Here we profile patients in...

Since the discovery of messenger RNA (mRNA), there have been tremendous efforts to wield them in the development of therapeutics and vaccines. During the COVID-19 pandemic, two mRNA vaccines were developed and approved in record-breaking time, revolutionizing the vaccine development landscape. Although first-generation COVID-19 mRNA vaccines have d...

Background: The impact of COVID-19 severity on development of long-term sequelae remains unclear, and symptom courses are not well defined. Methods: This ambidirectional cohort study recruited adults with new or worsening symptoms lasting ≥3 weeks from confirmed SARS-CoV-2 infection between August 2020-December 2021. COVID-19 severity was define...

Systemic Lupus Erythematosus (SLE) is an autoimmune disease characterized by multiple autoantibodies, some of which are present in high titers in a sustained, B cell-independent fashion consistent with their generation from long-lived plasma cells (LLPC). Active SLE displays high numbers of circulating antibody-secreting cells (ASC). Understanding...

Novel mRNA vaccines have resulted in a reduced number of SARS-CoV-2 infections and hospitalizations. Yet, there is a paucity of studies regarding their effectiveness on immunocompromised autoimmune subjects. In this study, we enrolled subjects naïve to SARS-CoV-2 infections from two cohorts of healthy donors (HD, n=56) and systemic lupus erythemato...

Novel mRNA vaccines have resulted in a reduced number of SARS-CoV-2 infections and hospitalizations. Yet, there is a paucity of studies regarding their effectiveness on immunocompromised autoimmune subjects. In this study, we enrolled subjects naive to SARS- CoV-2 infections from two cohorts of healthy donors (HD, n=56) and systemic lupus erythemat...

Immunoglobulin E (IgE) is central to the pathogenesis of allergic conditions including allergic fungal rhinosinusitis (AFRS). However, little is known about IgE antibody-secreting cells (ASCs). We performed single cell RNA sequencing (scRNA-seq) from CD19+ and CD19neg ASCs of nasal polyps (NPs) from patients with AFRS (n = 3). NPs were highly enric...

Severe SARS-CoV-2 infection1 has been associated with highly inflammatory immune activation since the earliest days of the COVID-19 pandemic2-5. More recently, these responses have been associated with the emergence of self-reactive antibodies with pathologic potential6-10, although their origins and resolution have remained unclear11. Previously,...

Infection with SARS‐CoV‐2, the etiology of the ongoing COVID‐19 pandemic, has resulted in over 450 million cases with more than 6 million deaths worldwide, causing global disruptions since early 2020. Memory B cells and durable antibody protection from long‐lived plasma cells (LLPC) are the mainstay of most effective vaccines. However, ending the p...

Following infection or vaccination, antibody-secreting cells (ASC), also called plasmablasts, appear in circulation transiently; a small fraction of them then migrate to the spleen or bone marrow (BM) to mature into long-lived plasma cells (LLPC). While LLPC, by definition, are quiescent or non-dividing, early-minted blood ASC were mostly thought t...

Early-minted antibody-secreting cells (ASC) undergo further maturation to become long-lived plasma cells (LLPC) in the bone marrow (BM) microniche to afford life-long humoral protection. As terminally-differentiated cells, ASC are thought to be antibody factories that secrete immunoglobulins (Ig) constitutively. This model suggests that Ig secretio...

Critical technological advances have enabled the rapid investigations into the immune responses elicited by SARS-CoV-2, the pathogen responsible for the COVID-19 pandemic. We discuss the cutting-edge methods used to deconvolve the B-cell responses against this virus and the impact they have had in the ongoing public health crisis.

Antibody secreting cells (ASCs) circulate after vaccination and infection and migrate to the BM where a subset known as long-lived plasma cells (LLPCs) persists and secrete antibodies for a lifetime. The mechanisms by which circulating ASCs become LLPCs are not well elucidated. Here, we show that human blood ASCs have distinct morphology, transcrip...

Recent studies have demonstrated the significant breadth of emerging autoreactivity in severe SARS-CoV-2 infection. Importantly, we have identified a relaxation of peripheral tolerance within early antibody secreting cells that emerge in patients with COVID-19 as important drivers of those responses. While often viral-specific, these extrafollicula...

Objective While the contribution of B-cells to SLE is well established, its role in chronic cutaneous lupus erythematosus (CCLE) remains unclear. Here, we compare B-cell and serum auto-antibody profiles between patients with systemic lupus erythematosus (SLE), CCLE, and overlap conditions. Methods B-cells were compared by flow cytometry amongst he...

Antibody secreting cells (ASC) circulate after vaccination and migrate to the bone marrow (BM) where a subset known as long-lived plasma cells (LLPC) persist and secrete antibodies for a lifetime. The mechanisms of how circulating ASC become LLPC are not well elucidated. Here, we show that human blood ASCs have distinct morphology, transcriptomes,...

Increased IgE is a typical feature of allergic rhinitis. Local class-switch recombination has been intimated but B cell precursors and mechanisms remain elusive. Here we describe the dynamics underlying the generation of IgE-antibody secreting cells (ASC) in human nasal polyps (NP), mucosal tissues rich in ASC without germinal centers (GC). Using V...

In late 2019, the emergence of the coronavirus SARS-CoV-2 resulted in a global pandemic, costing over 2 million lives. A hallmark of the disease resulting from SARS-CoV-2 infection, COVID-19, has been it’s heterogeneity in presentation. To understand the immunological underpinnings of this variability in symptoms, we performed a deep characterizati...

A wide spectrum of clinical manifestations has become a hallmark of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) COVID-19 pandemic, although the immunological underpinnings of diverse disease outcomes remain to be defined. We performed detailed characterization of B cell responses through high-dimensional flow cytometry to revea...

Severe SARS-CoV-2 infection is linked to the presence of autoantibodies against multiple targets, including phospholipids and type-I interferons. We recently identified activation of an autoimmune-prone B cell response pathway as correlate of severe COVID-19, raising the possibility of de novo autoreactive antibody production during the antiviral r...

Background: B cell affinity maturation enables B cells to generate high-affinity antibodies. This process involves somatic hypermutation of B cell immunoglobulin receptor (BCR) genes and selection by their ability to bind antigens. Lineage trees are used to describe this microevolution of B cell immunoglobulin genes. In a lineage tree, each node i...

The squalene-based oil-in-water emulsion (SE) vaccine adjuvant MF59 has been administered to more than 100 million people in more than 30 countries, in both seasonal and pandemic influenza vaccines. Despite its wide use and efficacy, its mechanisms of action remain unclear. In this study we demonstrate that immunization of mice with MF59 or its mim...

The squalene-based oil-in-water emulsion (SE) vaccine adjuvant MF59 has been administered to more than 100 million people in more than 30 countries, in both seasonal and pandemic influenza vaccines. Despite its wide use and efficacy, its mechanisms of action remain unclear. In this study we demonstrate that immunization of mice with MF59 or its mim...

Introduction A wide clinical spectrum has become a hallmark of the SARS-CoV-2 (COVID-19) pandemic, although its immunologic underpinnings remain to be defined. We have performed deep characterization of B cell responses through high-dimensional flow cytometry to reveal substantial heterogeneity in both effector and immature populations. More notabl...

Allergic fungal rhinosinusitis (AFRS) is a subset of chronic rhinosinusitis with nasal polyposis (CRSwNP). AFRS is primarily associated with a type 2 inflammatory profile including eosinophilic mucosal infiltration and high serum and mucosal titers of IgE. Patients with AFRS have increased frequencies of IgE ASCs within NP structures, although it i...

The immunoglobulin genes of germinal center B cells (GCB) undergo somatic hypermutation in order to generate antibody diversity. Additionally, several non-immunoglobulin (non-Ig) genes are known to accumulate mutations in the germinal center. There are two methods by which mutations are introduced at the Ig and non-Ig loci. The first is through the...

The maintenance of immunological tolerance of B lymphocytes is a complex and critical process that must be implemented as to avoid the detrimental development of autoreactivity and possible autoimmunity. Murine models have been invaluable to elucidate many of the key components in B‐cell tolerance; however, translation to human homeostatic and path...

The increasingly recognized role of different types of B cells and plasma cells in protective and pathogenic immune responses combined with technological advances have generated a plethora of information regarding the heterogeneity of this human immune compartment. Unfortunately, the lack of a consistent classification of human B cells also creates...

Chronic autoimmune diseases, and in particular Systemic Lupus Erythematosus (SLE), are endowed with a long‐standing autoreactive B‐cell compartment that is presumed to reactivate periodically leading to the generation of new bursts of pathogenic antibody‐secreting cells (ASC). Moreover, pathogenic autoantibodies are typically characterized by a hig...

B cell affinity maturation enables B cells to generate high-affinity antibodies. This process involves somatic hypermutation of B cell immunoglobulin receptor (BCR) genes and selection by their ability to bind antigens. Lineage trees are used to describe this microevolution of B cell immunoglobulin genes. In a lineage tree, each node is one BCR seq...

The immune system responds preferentially to particular antigenic-epitopes contained within complex immunogens, such as proteins or microbes. This poorly understood phenomenon, termed ‘‘immunodominance,’’ remains an obstacle to achieving polyvalent immune responses against multiple anti-genic-epitopes through vaccination. We observed profound suppr...

Systemic Lupus Erythematosus (SLE) is characterized by B cells lacking IgD and CD27 (double negative; DN). We show that DN cell expansions reflected a subset of CXCR5⁻ CD11c⁺ cells (DN2) representing pre-plasma cells (PC). DN2 cells predominated in African-American patients with active disease and nephritis, anti-Smith and anti-RNA autoantibodies....

Cancer-associated fibroblasts (CAFs) are generally associated with poor clinical outcome. CAFs support tumor growth in a variety of ways and can suppress antitumor immunity and response to immunotherapy. However, a precise understanding of CAF contributions to tumor growth and therapeutic response is lacking. Discrepancies in this field of study ma...

MF59 is an oil-in-water emulsion adjuvant approved for human influenza vaccination in European Union. The mode of action of MF59 is not fully elucidated yet, but results from several years of investigation indicate that MF59 establishes an immunocompetent environment at injection site which promotes recruitment of immune cells, including antigen pr...

Phenotypic characterization of APCs within the LN. Flow cytometry dot plots representing the gating strategy to define APCs within LN cell suspension of naïve mice are shown. Each reported dot plot derives from the previous one as indicated by the gray arrows and the numbers. Cell suspension of popliteal LNs was obtained and labeled as described in...

Mo-DCs accumulation within dLNs is observed also upon intramuscular immunization and occurred three days after immunization with ALUM adjuvant. (a) Mice were immunized between toes (b.t.) or intramuscularly (i.m.) with PE alone or adjuvanted with MF59. Cell suspensions of popliteal dLNs were analyzed by flow cytometry 18 hrs after the immunization....

APCs from dLNs of mice immunized with MF59 enhance the triggering of antigen-specific T cell response. (a) The bar graph histogram shows the percentage of CFSE halving (which measures the proliferating cells) of OT-II OVA-specific CD4 T cells cultured with APCs derived from dLNs of mice collected 18 hrs after immunization with with OVA (white bar)...

Real time translocation to popliteal dLN of PE injected between toes. Mice were injected b.t. with PE and a real time intravital imaging analysis of the popliteal dLN was taken 15 minutes from the injection (procedure described in Materials and Methods section). The time-lapse of the antigen translocation to popliteal dLN is reported. In red is sho...

3D antigen and MF59 localization within the dLN: 18 hrs after injection the antigen is preferentially, whereas MF59 is totally, accumulated within the medullary compartment of the LN. Mice were immunized b.t. with PE and fluorescently labelled-MF59 and popliteal dLNs were collected, processed and analyzed by two-photon microscopy (all the procedure...

Phenotype of intranodal differentiated Mo-DCs. (a) Flow cytometry histograms of CD11c expression by the CD8α_F4/80 double positive APC subset which is CD11c negative/low 8 hrs (grey line) but positive 18 hrs (red line) after immunization with MF59. The CD11c MFI of the CD8α_F4/80 APCs at 8 hrs and 18 hrs from the treatment is also reported. (b) The...

Only Mo-DCs display up-regulation of CD86 co-stimulatory molecule. Flow cytometry histograms of CD86 expression by MΦs (blue), DCs (green) and Mo-DCs (red) from mice immunized with PE (thin lines) or PE + MF59 (thick lines). The MFI of CD86 expression is reported per each APC subset. Representative results of one experiment out of two are reported....

MF59 works as adjuvant using PE as model antigen. Graph reports the anti-PE IgG titers in the mouse sera collected two weeks after the second immunization with PE (white dots) or PE + MF59 (black dots) in two independent experiments, as indicated (exp. 1; exp. 2). Each dot depicts the antibody titer from a single mouse. The arithmetic mean of the v...

3D antigen localization within the dLN: 18 hrs after injection the antigen is preferentially accumulated within the medullary compartment of the LN. Mice were immunized b.t. with PE and popliteal dLNs were collected, processed and analyzed by two-photon microscopy (all the procedures are described in Materials and Methods section). 3D-imaging of th...

Phenotype of monocytes versus Mo-DCs reveals that Mo-DCs accumulate within the dLN only 18 hrs after the immunization with MF59. (a) Flow cytometry dot plots representing the expression of CD8α and CD64 (upper quadrant) or CD8α and CD11c (lower quadrant) in APCs, 8 hrs after the immunization with PE alone. The few CD8α_F4/80 double positive APCs (r...

Expression of MCH class II molecules by APCs in MF59 immunized mice. Flow cytometry dot plots reporting the profile of MHC class II expression by MΦs (blue, left panel), Mo-DCs (red, central panel) and DCs (green, right panel) after 18 hrs from the immunization with PE + MF59. Representative results of one experiment out of two are shown. (TIF)

MF59 promotes the persistence of antigen-loaded APCs within the dLN during the first 18 hours. (a) Flow cytometry dot plots of PE-positive dLN cells (left dot plot) and PE-positive APCs (central and right dot plots) 18 hrs after immunization, identified according to the gating strategy reported in S1 Fig, are shown. The majority of PE-loaded dLN ce...

Mo-DCs are mostly double loaded with PE and MF59 compared to DCs and MΦs. Graph bar histogram report the average percentage (+ standard deviation) of MΦs (blue bar), DCs (green bar) and Mo-DCs (red bar) double positive for PE and MF59 (PE/MF59) 18 hours after the immunization. Results of two independent experiments are plotted. Statistical analysis...

Real time translocation to popliteal dLN of PE and MF59 injected between toes. Mice were injected b.t. with PE + fluorescently labelled-MF59 and a real time intravital imaging analysis of the popliteal dLN was taken 1 hour from the injection (procedure described in Materials and Methods section). The time-lapse of the antigen translocation to popli...

Despite the reported clinical benefits of checkpoint blockade in some cancers, a large proportion of patients fail to respond, and insights from preclinical and clinical studies have indicated that therapy failure may stem from cues emanating from the tumor microenvironment. Strikingly, resistance to PD-1 immunotherapy in patients with metastatic m...

A previous study found that NF-κB activation is delayed in L929 cells infected with wild-type (wt) strains of VSV, while activation occurred earlier in cells infected with mutant strain T1026R1 (R1) that encodes a mutation in the cytotoxic matrix (M) protein. The integrity of the other R1 proteins is unknown; therefore our goal was to identify the...

Background: Lymph nodes (LNs) are positioned strategically throughout the body as critical mediators of lymph filtration and immune response. Lymph carries cytokines, antigens, and cells to the downstream LNs, and their effective delivery to the correct location within the LN directly impacts the quality and quantity of immune response. Despite th...

In lymph nodes, fibroblastic reticular cells (FRCs) form a collagen-based reticular network that supports migratory dendritic cells (DCs) and T cells and transports lymph. A hallmark of FRCs is their propensity to contract collagen, yet this function is poorly understood. Here we demonstrate that podoplanin (PDPN) regulates actomyosin contractility...

Fibroblastic reticular cells (FRCs) are known to inhabit T cell-rich areas of lymphoid organs, where they function to facilitate interactions between T cells and dendritic cells. However, in vivo manipulation of FRCs has been limited by a dearth of genetic tools that target this lineage. Here, using a mouse model to conditionally ablate FRCs, we de...

Dendritic cells (DCs) are well established as potent antigen-presenting cells critical to adaptive immunity. In vaccination approaches, appropriately stimulating lymph node-resident DCs (LNDCs) is highly relevant to effective immunization. Although LNDCs have been implicated in immune response, their ability to directly drive effective immunity to...

Migrating dendritic cells follow precise navigational chemokine gradients established by lymph node stromal cells through their asymmetric expression of the atypical chemokine receptor CCRL1.

Papain, a cysteine protease allergen with inherent adjuvant activity, induces potent IL-4 expression by T cells in the popliteal lymph nodes of mice following footpad immunization. In this study, we identify a novel, non-BCR-mediated capacity for B cells to rapidly bind and internalize papain. B cells subsequently regulate the adaptive immune respo...

Many common allergens such as house dust mite antigen and grass pollen are complex mixes of proteins and other compounds, both of which may have innate signaling capacity. Understanding the role of each of these signals is essential to determining what separates allergens from other innocuous proteins. Papain is a cysteine protease structurally sim...

Background: As fluorescent microscopy has developed, significant insights have been gained into the establishment of immune response within secondary lymphoid organs, particularly in draining lymph nodes. While established techniques such as confocal imaging and intravital multi-photon microscopy have proven invaluable, they provide limited insigh...

The DCs were labeled with cell-tracker dyes; WT DCs are shown in red and Clec1b−/− DCs are shown in green. Frames are 90 s apart and shown at 8 frames/s.

Frames are 2 min apart and shown at 8 frames/s.

Control DCs are shown first, followed by DCs treated with rhodocytin and PDPN-Fc. Frames are 90 s apart and shown at 8 frames/s. Scale bar represents 20 μm.

Frames are 2 min apart and shown at 8 frames/s.

CFSE-labeled DCs (green), second harmonic (collagen fibers) (red). Frames are 30 s apart and shown at 8 frames/s.

Frames are 90 s apart and shown at 8 frames/s.