Meditation-induced bloodborne factors as an adjuvant treatment to COVID-19 disease

Abstract

The COVID-19 pandemic has resulted in significant morbidity and mortality worldwide. Management of the pandemic has relied mainly on SARS-CoV-2 vaccines, while alternative approaches such as meditation, shown to improve immunity, have been largely unexplored. Here, we probe the relationship between meditation and COVID-19 disease and directly test the impact of meditation on the induction of a blood environment that modulates viral infection. We found a significant inverse correlation between length of meditation practice and SARS-CoV-2 infection as well as accelerated resolution of symptomology of those infected. A meditation "dosing" effect was also observed. In cultured human lung cells, blood from experienced meditators induced factors that prevented entry of pseudotyped viruses for SARS-CoV-2 spike protein of both the wild-type Wuhan-1 virus and the Delta variant. We identified and validated SERPINA5, a serine protease inhibitor, as one possible protein factor in the blood of meditators that is necessary and sufficient for limiting pseudovirus entry into cells. In summary, we conclude that meditation can enhance resiliency to viral infection and may serve as a possible adjuvant therapy in the management of the COVID-19 pandemic.

Keywords: Adoptive blood transfer; COVID-19 disease; Immunity; Meditation; Pseudovirus; SARS-CoV-2; serpin.

Fig. 1

Graphical representation of meditator survey data. a-d, Among lowest-resolution geospatial communities meeting a minimum sample size of 10, relationships between the proportion of participants from a 7-day meditation workshop having at least six months of meditation experience and their self-reported rates of COVID-19 infection (a), brain fog (b), congestion (c), and duration of illness (d). e, Relationship between the proportion of participants from a 7-day meditation workshop, grouped by geospatial area, that meditates daily and rate of anosmia. f, Among individual respondents, the relationship between meditation frequency and probability of COVID-19 infection. g-i, Among individual respondents, the relationship between meditation experience (binary; six months of experience) and COVID-19 infection (g), brain fog (h), and duration of illness (i). j-l, Among individual respondents, the relationship of multivariable predictors selected by backwards selection (Wald χ2) and COVID-19 infection (j), brain fog (k), and duration of illness (l). m, Among individual respondents, relationship between the probability of COVID-19 and vaccination status. n, Among individual respondents, the relationship of multivariable predictors and COVID-19 infection. All regression curves were plotted with LOESS (span α = 1.50).

Fig. 2

Meditation induces the expression of bloodborne factors that protect against SARS-CoV-2 infection. a, Schematic representation of SARS-CoV-2.SD19-RFP pseudovirus construction and transmission electron microscopy (TEM) image of virion. b, Human lung cells (A-549) and mouse muscle cells (C2C12) 24 h after exposure to SARS-CoV-2.SD19-RFP. c, Representative immunofluorescence images of A549 cells 24 h after exposure to pseudovirus. Negative control represents vehicle only while positive control cells were exposed to pseudovirus (upper panels). Lower panels represent cells pre-treated with either experienced pre- (left panel) or post- (right panel) meditation plasma prior to addition of pseudovirus. Red, red fluorescent protein (RFP); Blue, Hoechst 33342. Scale bars, 50 μm d, Scanning EM (SEM) (upper and middle panels) and TEM (lower panels) images of A549 cells treated with post-meditation plasma from control, novice, or experienced meditators, followed by addition of pseudovirus. e, Quantification of red fluorescence intensity in A549 cells treated with control, novice, or experienced pre- and post-meditation plasma followed by addition of pseudovirus. Two-tailed analysis, Mann-Whitney post-test (control n= 21, novice n= 45, experienced n= 43) was performed (*p < 0.05). f-g, Correlation of sex with red fluorescence intensity in A549 cells treated with control, novice, or experienced pre- and post-meditation male (f) and female (g) plasma followed by addition of pseudovirus. Two-tailed analysis, Mann-Whitney post-test (control n= 8 male, 13 female; novice n= 23 male, 22 female; experienced n= 17 male, 26 female) was performed (*p < 0.05). h, Pre-post correlation of red fluorescence intensity with age in A549 cells treated with control, novice, or experienced meditation plasma. A positive pre-post value is indicative of a positive correlation of age and protection from pseudoviral infectivity, while a negative pre-post value demonstrates a negative correlation. Two-tailed analysis, Mann-Whitney post-test (control n= 20, novice n= 44, experienced n= 43) was performed (*p < 0.05). i-k, A549 cells were pre-treated with heat-inactivated (i) or ultra-centrifuged plasma supernatant (j) or pellet (k) from control, novice, or experienced meditators pre- and post-meditation, followed by addition of pseudovirus. Cells were fixed and stained 24 h after addition of pseudovirus. Two-tailed analysis, Mann-Whitney post-test was performed (*p < 0.05). Blue dots, pre-meditation; red dots, post-meditation. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Fig. 3

Experienced meditators have elevated levels of plasma SERPINA5. a, Pseudovirus interactome co-immunoprecipitated from control and experienced post-meditation plasma using spike antibody. b, SERPINA5 concentration (pg/mL) in control and experienced post-meditation plasma (control n=18, experienced n=40). c, Correlation analysis between plasma SERPINA5 concentration (pg/mL) and viral infection protection (RFP/DAPI) in novice (n = 37, gray dots) and experienced (n = 33, blue dots) meditators. r, Pearson's correlation value, (*p < 0.05). d, Ribbon diagram of SERPINA1 (3CWM) and SERPINA5 (2OL2) superimposed on one another. SERPINA1 and SERPINA5 were aligned with the RCSB pairwise structure alignment tool (Berman et al., 2000) using the iFACTCAT(rigid) algorithm (Li et al., 2020). The reactive center loop (RCL) is depicted in the right panel for all inhibitory clade A SERPINs. The top alignment represents color by amino acid while the bottom alignment represents similarity as calculated by Blosum62 score matrix in Geneious (G. 11.1.5). Green amino acids represent 80–100% similarity, yellow amino acids represent 60–80% similarity, and orange amino acids represent less than 60% similarity. Both N- and C-terminal exocites are intact as well as a central Ser (P1|P1’) with the exception of SERPINA12. Alignment scheme adapted from Sanrattana et al. (Sanrattana et al., 2019). e, Ribbon diagrams of TMPRSS2 (PDB 7MEQ) (catalytic domain, depicted in red; left panel), SERPINA5 (2OL2) with RCL (depicted in red; middle panel). Beta sheets are shown in green, α-helices in blue. Potential TMPRSS2/SERPINA5 docking structures are indicated (right panel). Firedock global energy (3.21), attractive VdW (−1.35), Repulsive VdW (0.49), ACE (−0.46), HB (0.0). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Fig. 4

Effect of post-meditation plasma, vaccination, and SERPINA5 on infectivity of SARS-CoV-2 virus variants. a-f (upper panels), Effect of plasma from control, novice, and experienced meditators on SARS-CoV-2 pseudovirus variant infectivity in A549 cells with Delta (a), Lambda (b), Beta (c), U.K. (d), Brazil (e), or D614G (f) variants. Viral constructs were added to cultured cells following pre-treatment with control or meditation plasma. Two-tailed analysis, Mann-Whitney post-test (control n= 23, novice n= 45, experienced n= 43) was performed (*p < 0.05). (lower panels), Correlation analysis between plasma SERPINA5 concentration (pg/mL) and viral infection protection (RFP/DAPI) in novice (n = 37, gray dots) and experienced (n = 33, blue dots) meditators. r, Pearson's correlation value, (*p < 0.05). g, Effect of post-vaccination plasma on SARS-CoV-2 pseudovirus variant infectivity in A549 cells. Two-tailed analysis, Mann-Whitney post-test was performed (*p < 0.05). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)