Very excited to share our latest research on immunological features of #LongCovid. Our 2+ year collaboration with @PutrinoLab with many other fantastic colleagues and patients - Mount Sinai Yale Long COVID (MY-LC) study by @sneakyvirus1 et al. 🧵(1/)
There are multiple hypotheses behind long COVID pathogenesis including persistent virus/viral remnants, autoimmunity, dysbiosis, virome reactivation and tissue damage. Our data will dive deep into some of these. (3/)
This is a cross-sectional multi-dimensional immune phenotyping & patient surveys in people with or without LC, who got COVID during the 2020 first wave, on average more than a year from infection. Most were not-hospitalized, ♀ dominant, younger to middle age. (4/)
Long COVID group reported significant increases in the intensity of symptoms and dramatically worsened quality of life. Survey outcomes put together into a single classification metric “Long COVID Propensity Score or LCPS” demonstrated significant diagnostic potential. (5/)
Long COVID participants reported a number of symptoms, most commonly fatigue, brain fog, dysautonomia..etc. Hierarchical clustering of binary symptom data identified 3 clusters of patients with similar sets of self-reported symptoms. (6/)
Next, flow cytometry analysis @peowenlu of peripheral blood mononuclear cells revealed several key differences in LC vs. CC or HC. First, LC had increase in non-conventional monocytes, activated B cells double-negative B cells, and decrease in conventional dendritic cells 1. (7/)
Long COVID participants also had reduced central memory T cells and increased exhausted CD4 and CD8 T cells. The exhausted T cells suggest chronic antigens stimulating these T cells. What they are we do not know yet. (8/)
Long COVID patients also had increases in CD4 T cells that secrete IL-2, IL-4 and IL-6, as well as some that secrete both IL-4 and IL-6. These T cells correlated with the levels of EBV reactive antibodies. Follow me down this thread further to find out more! (9/)
We measured antibody levels against SARS-CoV-2 antigens in people who received 2 doses of mRNA vaccines. Curiously, long COVID patients produced higher levels of IgG against Spike. Without vaccines, LC had higher IgG against nucleocapsid. Data suggest persistent antigen? (10/)
Not only did LC have higher levels of IgG against Spike, but also had IgG against distinct epitopes within the Spike protein, identified by @S_Tabachnikova with @serimmune Kathy Kamath. See the distinct peaks in purple vs. controls. (11/)
Next we examined a large number of plasma factors and asked which factors are most different in long COVID vs. non-long COVID groups. By far the most significant differences were found in cortisol levels. Long COVID group had lower plasma cortisol levels than control groups.(12/)
Cortisol levels in circulation were about half of the control groups. Despite this, we saw no elevation in ACTH levels, suggesting an impaired compensatory response by the hypothalamic-pituitary axis. (13/)
This is so interesting, giving the report by @SuYapeng et al showing similar reduction in long haulers with respiratory viral symptoms at 2-3 months post COVID. This implies chronic hypocortisolism in long COVID. (14/)
What about autoantibodies? @_BlueJay3 and @aaronmring used Rapid Extracellular Antigen Profiling (REAP) version 2.0 (with more antigens added) to examine autoantibodies. In contrast to what we found for acute severe COVID, long COVID group did not have elevated AABs. (15/)
However, REAP did find notably elevated autoantibodies to sodium ion transporters in a subset of Long COVID patients who displayed reactivities agains 6-9 different proteins of this family. Those with tinnitus and nausea had elevated levels of these AABs. (16/)
In contrast to autoantibodies, REAP detected elevation in IgG against herpesvirus antigens. In particular, antibody reactivity to glycoproteins and early antigens of Epstein-Barr virus, Varicella zoster virus were elevated in long COVID over other groups. (17/)
The increases in antibodies to EBV and VZV antigens were also detected using independent assays like ELISA and @serimmune epitope mapping. However, seroprevalence for EBV and VZV were similar in LC and CC. These data suggest recent reactivation of EBV and VZV in LC. (18/)
This again is consistent with the report by @SuYapeng et al, showing that EBV viremia at the time of diagnosis is one of the four predictive factors for long COVID. (Note that our study did not examine viremia but infer EBV reactivation by serology)(19/)
Notably, these anti-EBV antibodies found in LC correlated with the levels of IL-4/IL-6 double-positive CD4 T cells I mentioned above. Significant correlations were also found between EBV p23 reactivity and terminally differentiated effector memory (TEMRA) CD4 T. (20/)
Finally @rahuldhodapkar used machine learning and found that immune features alone can predict long COVID with efficient discriminative performance (AUC=0.96)! The most informative individual data blocks contributing to efficient separation of groups are flow and cytokine. (21/)
Several features significantly distinguished Long COVID (double negative B cells, serum galectin-1, various EBV epitopes) while others were negatively associated (serum cortisol, PD-1+ CD4+ TCM, and HSV1 and HSV2 motifs). (22/)
In fact, serum cortisol was the most significant individual predictor of Long COVID status in the model, and alone was a predictor wit an AUC of 0.96 (95% CI: 0.92-0.99). Notably, serum cortisol within the MY-LC study was highest in HC > CC > LC. (23/)
We found many key circulating biological factors that alone can discriminate long COVID from others. Comparison of classification accuracies between patient reported outcomes and machine learning revealed substantial agreement (Cohen’s Kappa .865, 95% CI [0.83 - .90]). (24/)
While we are so excited about the findings of this study, we also wish to highlight key limitations of this study. They are summarized here but there may be more. Our study is exploratory in nature and requires validation. (25/)
Here is the summary of our findings. There are many implications for diagnosis and therapy for #longCOVID. (26/)
For example, can we target viral reservoir with antivirals and mAbs? Can we restore cortisol levels? Should therapy be targeting EBV? Would immunomodulatory therapy against inflammatory cytokines be useful? We still need to identify long COVID endotypes for treatment. (27/)
We hope that our study will be informative to others working in the field. We need validation across cohorts. We also hope that these data will help those who are still skeptical understand that long COVID is real, and it has biological basis. Thank you for reading. (End)
And I finally got to meet @PutrinoLab in person after a million zoom meetings and phone calls. How amazing to have a collaborator who is also a great friend!! 😊
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A new study led by @marioph13 in collaboration with the @WilenLab examines two bat coronaviruses that are the closest relatives of SARS-CoV-2 for their ability to infect, evade immunity and transmit between rodents. Some key takeaway points 🧵 (1/) nature.com/articles/s4156…
Here is a link to the accessible manuscript (2/) rdcu.be/dPjsA
We used the highest biosafety level available at Yale (BSL3+) to conduct the study and we did not introduce gain-of-function mutations into the bat viruses. No serial passaging of the viruses were done - to avoid adaptation. Grateful to @YaleEHS for all the support. (3/)
Sharing our new study by @keylas3, @SilvaJ_C, Rafael Bayarri Olmos et al (with T. Horvath & @PutrinoLab) showing that a passive transfer of IgG from patients with #longCOVID into mice recapitulates ⬆️ pain and other symptoms 🧵 (1/)
Long COVID disease pathogenesis includes persistent SARS-CoV-2 virus, dysbiosis, herpesvirus reactivation, autoimmunity, and others. In this study, we focus on the role of autoantibodies. (2/)
Among the original Mount Sinai-Yale Long COVID study participants 👇🏼 (with @PutrinoLab), we focused on patients with high neurological symptom burden (n=55), and compared antibodies with convalescent controls (n=42) or uninfected controls (n=39). (3/) nature.com/articles/s4158…
What determines whether someone gets infected or not after exposure to SARS-CoV-2? A new study by Lindeboom et al examined this question with COVID-19 human challenge study. @BenIsraelow and I summarize their key findings in this News & Views 🧵 (1/) nature.com/articles/d4158…
The study: 16 healthy young volunteers with no prior infection or vaccination were inoculated nasally with a low dose of pre-Alpha SARS-CoV-2 strain. Interestingly, only 6 had sustained infection, 3 had transient, and 7 had abortive infection at this dose. (2/) nature.com/articles/s4158…
The three infection outcomes allowed investigation of key features associated with susceptibility vs. resistance. Higher baseline expression of HLA-QA2 mRNA was associated with COVID resistance. Early nasal interferon I response was seen in those with transient infection. (3/)
Preventing infection is the best way to avoid diseases like #PAIS. A new study from our team @tianyangmao, Jooyoung Kim, @marioph13 et al shows that a generic antibiotic neomycin acts on the host immune system in the👃🏽to trigger antiviral resistance. (1/)🧵 pnas.org/doi/10.1073/pn…
This work is inspired by @SmitaGopinath et al who showed that an antibiotic class called aminoglycosides has an unusual antiviral property. Aminoglycosides including neomycin trigger interferon-stimulated genes through a TLR3-dependent mechanism. (2/) ncbi.nlm.nih.gov/pmc/articles/P…
In our current study, we showed that nasal application of neomycin in mice one day before infection reduces viral load and disease burden after the SARS-COV-2 challenge. @tianyangmao (3/)
Delighted to share our latest work on #longCOVID - sex differences in symptoms and immune signatures. Led by @SilvaJ_C @taka_takehiro @wood_jamie_1 et al. with @LeyingGuan & @PutrinoLab. We find a striking inverse correlation btw testosterone levels and symptom burden👇🏼 (1/)
This work leverages data from our recent Mount Sinai-Yale long COVID "MY-LC" study with the @PutrinoLab. This time, we asked the question, "Are there differences in symptoms and immune signatures of ♀️ vs. ♂️ with LC"? (2/)
While some symptoms were equally frequent in females and males, many were more frequent in females (e.g., swelling, headaches, muscle pain, cramps) than males. The top distinguishing symptoms of LC status by sex were hair loss in females and sexual dysfunction in males. (3/)
In this prospective observational study, we examined changes in symptoms & immune phenotypes in vaccine-naïve people with #LongCovid after COVID-19 vaccination. Due to the timing of the initiation of this study, we were only able to recruit 16 people. However, the insights we gained are intriguing. Led by @connorbgrady, @bornali_27, @silva_JC, @hmkyale et al. (1/) medrxiv.org/content/10.110…
This study was initiated in collaboration with @Survivor_Corps @dianaberrent based on their Facebook poll showing that 40% of respondents with self-reported Long COVID had mild to full symptom resolution after vaccination while 14% reported worsening of their symptoms. (2/) doi.org/10.1101/2021.0…
In addition, evidence from other patient advocate groups, including @patientled and @longCovidSOS, and from @DanielGriffinMD, was emerging at the time on the impact of COVID vaccines in people with long COVID. (3/)