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Aging-dependent alterations in gene expression and a mitochondrial signature of responsiveness to human influenza vaccination.

Thakar J, Mohanty S, West AP, Joshi SR, Ueda I, Wilson J, Meng H, Blevins TP, Tsang S, Trentalange M, Siconolfi B, Park K, Gill TM, Belshe RB, Kaech SM, Shadel GS, Kleinstein SH, Shaw AC - Aging (Albany NY) (2015)

Bottom Line: To elucidate gene expression pathways underlying age-associated impairment in influenza vaccine response, we screened young (age 21-30) and older (age≥65) adults receiving influenza vaccine in two consecutive seasons and identified those with strong or absent response to vaccine, including a subset of older adults meeting criteria for frailty.The response signature was dysregulated in older adults, with the plasma cell signature induced at day 2, and was never induced in frail subjects (who were all non-responders).These results represent the first genome-wide transcriptional profiling analysis of age-associated dynamics following influenza vaccination, and implicate changes in mitochondrial biogenesis and function as a critical factor in human vaccine responsiveness.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Yale School of Medicine, New Haven, CT 06520, USA.

ABSTRACT
To elucidate gene expression pathways underlying age-associated impairment in influenza vaccine response, we screened young (age 21-30) and older (age≥65) adults receiving influenza vaccine in two consecutive seasons and identified those with strong or absent response to vaccine, including a subset of older adults meeting criteria for frailty. PBMCs obtained prior to vaccination (Day 0) and at day 2 or 4, day 7 and day 28 post-vaccine were subjected to gene expression microarray analysis. We defined a response signature and also detected induction of a type I interferon response at day 2 and a plasma cell signature at day 7 post-vaccine in young responders. The response signature was dysregulated in older adults, with the plasma cell signature induced at day 2, and was never induced in frail subjects (who were all non-responders). We also identified a mitochondrial signature in young vaccine responders containing genes mediating mitochondrial biogenesis and oxidative phosphorylation that was consistent in two different vaccine seasons and verified by analyses of mitochondrial content and protein expression. These results represent the first genome-wide transcriptional profiling analysis of age-associated dynamics following influenza vaccination, and implicate changes in mitochondrial biogenesis and function as a critical factor in human vaccine responsiveness.

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Mitochondrial DNA and OXPHOS protein abundance confirm the elevated OXPHOS signature of responders(A) Relative mtDNA abundance in young (left panel) and older subjects (right panel) is significantly altered between vaccine responders (open boxes) and non-responders (filled boxes) at day 7 for young responders, with a trend at day 2 for older responders, compared to non-responders. mtDNA values are normalized with respect to the pre-vaccination time point. A subset from the cohort of older responders (n=7), older non-responders (n=8), young responders (n=6) and young non-responders (n=5) was evaluated. Dashed lines indicate the lower and upper range of values, with outliers indicated by open circles, bars indicating the 25th and 75th percentiles and dark horizontal lines the medians ** p<0.05, *p<0.1 (B) Induction of the mitochondrial chaperone protein HSP60 and mitochondrial Complex II component SDHA after influenza vaccination in young (left panel) compared to older adults (right panel), stratified by vaccine status (n=5 for each of the categories of older responder, older non-responder, young responder and young non-responder; relative pixel intensities of HSP60 and SDHA using β-actin as a loading control are normalized to day 0). For HSP60, a statistically significant increase in protein expression was found for young responders compared to non-responders at day 7 (p=0.04), and for older responders compared to non-responders at days 2 (p=0.007), 7 (p=0.0001), and 28 (p=0.0001). For SDHA, a significant increase in expression in young responders compared to non-responders was found at day 2 (p=0.007), and in older responders vs. older non-responders at days 2 (p=0.006), 7 (p=0.002), and 28 (p=0.007; p values were calculated using a t-test). (C) Representative Western blot of HSP60 and SDHA expression for young and older vaccine responders and non-responders.
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Figure 6: Mitochondrial DNA and OXPHOS protein abundance confirm the elevated OXPHOS signature of responders(A) Relative mtDNA abundance in young (left panel) and older subjects (right panel) is significantly altered between vaccine responders (open boxes) and non-responders (filled boxes) at day 7 for young responders, with a trend at day 2 for older responders, compared to non-responders. mtDNA values are normalized with respect to the pre-vaccination time point. A subset from the cohort of older responders (n=7), older non-responders (n=8), young responders (n=6) and young non-responders (n=5) was evaluated. Dashed lines indicate the lower and upper range of values, with outliers indicated by open circles, bars indicating the 25th and 75th percentiles and dark horizontal lines the medians ** p<0.05, *p<0.1 (B) Induction of the mitochondrial chaperone protein HSP60 and mitochondrial Complex II component SDHA after influenza vaccination in young (left panel) compared to older adults (right panel), stratified by vaccine status (n=5 for each of the categories of older responder, older non-responder, young responder and young non-responder; relative pixel intensities of HSP60 and SDHA using β-actin as a loading control are normalized to day 0). For HSP60, a statistically significant increase in protein expression was found for young responders compared to non-responders at day 7 (p=0.04), and for older responders compared to non-responders at days 2 (p=0.007), 7 (p=0.0001), and 28 (p=0.0001). For SDHA, a significant increase in expression in young responders compared to non-responders was found at day 2 (p=0.007), and in older responders vs. older non-responders at days 2 (p=0.006), 7 (p=0.002), and 28 (p=0.007; p values were calculated using a t-test). (C) Representative Western blot of HSP60 and SDHA expression for young and older vaccine responders and non-responders.

Mentions: To experimentally validate the OXPHOS gene expression signature of the responders and determine if this was associated with increased mitochondrial biogenesis (i.e. more mitochondria/cell), we examined mitochondrial DNA (mtDNA), which encodes 13 essential proteins of the OXPHOS chain and exists in multiple copies that usually correlate with organelle amount [14, 15]. Real-time qPCR analysis of mtDNA content in a subset of our cohort showed a significant increase in mtDNA copy number of young responders compared with young non-responders at 7 days post-vaccination (Figure 6A). In addition, older responders displayed elevated mtDNA copy number at day 2 post-vaccination, correlating with induction of the OXPHOS gene signature in this group (Figure 6A). We subsequently found that expression of HSP60, a mitochondrial matrix chaperone, was significantly elevated in protein extracts analyzed in a subset of responder PBMCs, compared to non-responders (at day 7 post-vaccination in young subjects and on all days in older subjects). In addition, mitochondrial succinate dehydrogenase subunit A (SDHA), a subunit of mitochondrial OXPHOS complex II, was also significantly elevated at the protein level in young responders vs. young non-responders at day 2 after vaccination, as well as in older responders vs. older non-responders at day 2, 7 and 28 (Figure 6B and 6C). This correlated with SDHA and many other OXPHOS genes being elevated based on transcriptional profiling. Taken together, these data are consistent with a role for mitochondrial biogenesis, perhaps driving increased TCA cycle and OXPHOS activity, in vaccine responders.


Aging-dependent alterations in gene expression and a mitochondrial signature of responsiveness to human influenza vaccination.

Thakar J, Mohanty S, West AP, Joshi SR, Ueda I, Wilson J, Meng H, Blevins TP, Tsang S, Trentalange M, Siconolfi B, Park K, Gill TM, Belshe RB, Kaech SM, Shadel GS, Kleinstein SH, Shaw AC - Aging (Albany NY) (2015)

Mitochondrial DNA and OXPHOS protein abundance confirm the elevated OXPHOS signature of responders(A) Relative mtDNA abundance in young (left panel) and older subjects (right panel) is significantly altered between vaccine responders (open boxes) and non-responders (filled boxes) at day 7 for young responders, with a trend at day 2 for older responders, compared to non-responders. mtDNA values are normalized with respect to the pre-vaccination time point. A subset from the cohort of older responders (n=7), older non-responders (n=8), young responders (n=6) and young non-responders (n=5) was evaluated. Dashed lines indicate the lower and upper range of values, with outliers indicated by open circles, bars indicating the 25th and 75th percentiles and dark horizontal lines the medians ** p<0.05, *p<0.1 (B) Induction of the mitochondrial chaperone protein HSP60 and mitochondrial Complex II component SDHA after influenza vaccination in young (left panel) compared to older adults (right panel), stratified by vaccine status (n=5 for each of the categories of older responder, older non-responder, young responder and young non-responder; relative pixel intensities of HSP60 and SDHA using β-actin as a loading control are normalized to day 0). For HSP60, a statistically significant increase in protein expression was found for young responders compared to non-responders at day 7 (p=0.04), and for older responders compared to non-responders at days 2 (p=0.007), 7 (p=0.0001), and 28 (p=0.0001). For SDHA, a significant increase in expression in young responders compared to non-responders was found at day 2 (p=0.007), and in older responders vs. older non-responders at days 2 (p=0.006), 7 (p=0.002), and 28 (p=0.007; p values were calculated using a t-test). (C) Representative Western blot of HSP60 and SDHA expression for young and older vaccine responders and non-responders.
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Figure 6: Mitochondrial DNA and OXPHOS protein abundance confirm the elevated OXPHOS signature of responders(A) Relative mtDNA abundance in young (left panel) and older subjects (right panel) is significantly altered between vaccine responders (open boxes) and non-responders (filled boxes) at day 7 for young responders, with a trend at day 2 for older responders, compared to non-responders. mtDNA values are normalized with respect to the pre-vaccination time point. A subset from the cohort of older responders (n=7), older non-responders (n=8), young responders (n=6) and young non-responders (n=5) was evaluated. Dashed lines indicate the lower and upper range of values, with outliers indicated by open circles, bars indicating the 25th and 75th percentiles and dark horizontal lines the medians ** p<0.05, *p<0.1 (B) Induction of the mitochondrial chaperone protein HSP60 and mitochondrial Complex II component SDHA after influenza vaccination in young (left panel) compared to older adults (right panel), stratified by vaccine status (n=5 for each of the categories of older responder, older non-responder, young responder and young non-responder; relative pixel intensities of HSP60 and SDHA using β-actin as a loading control are normalized to day 0). For HSP60, a statistically significant increase in protein expression was found for young responders compared to non-responders at day 7 (p=0.04), and for older responders compared to non-responders at days 2 (p=0.007), 7 (p=0.0001), and 28 (p=0.0001). For SDHA, a significant increase in expression in young responders compared to non-responders was found at day 2 (p=0.007), and in older responders vs. older non-responders at days 2 (p=0.006), 7 (p=0.002), and 28 (p=0.007; p values were calculated using a t-test). (C) Representative Western blot of HSP60 and SDHA expression for young and older vaccine responders and non-responders.
Mentions: To experimentally validate the OXPHOS gene expression signature of the responders and determine if this was associated with increased mitochondrial biogenesis (i.e. more mitochondria/cell), we examined mitochondrial DNA (mtDNA), which encodes 13 essential proteins of the OXPHOS chain and exists in multiple copies that usually correlate with organelle amount [14, 15]. Real-time qPCR analysis of mtDNA content in a subset of our cohort showed a significant increase in mtDNA copy number of young responders compared with young non-responders at 7 days post-vaccination (Figure 6A). In addition, older responders displayed elevated mtDNA copy number at day 2 post-vaccination, correlating with induction of the OXPHOS gene signature in this group (Figure 6A). We subsequently found that expression of HSP60, a mitochondrial matrix chaperone, was significantly elevated in protein extracts analyzed in a subset of responder PBMCs, compared to non-responders (at day 7 post-vaccination in young subjects and on all days in older subjects). In addition, mitochondrial succinate dehydrogenase subunit A (SDHA), a subunit of mitochondrial OXPHOS complex II, was also significantly elevated at the protein level in young responders vs. young non-responders at day 2 after vaccination, as well as in older responders vs. older non-responders at day 2, 7 and 28 (Figure 6B and 6C). This correlated with SDHA and many other OXPHOS genes being elevated based on transcriptional profiling. Taken together, these data are consistent with a role for mitochondrial biogenesis, perhaps driving increased TCA cycle and OXPHOS activity, in vaccine responders.

Bottom Line: To elucidate gene expression pathways underlying age-associated impairment in influenza vaccine response, we screened young (age 21-30) and older (age≥65) adults receiving influenza vaccine in two consecutive seasons and identified those with strong or absent response to vaccine, including a subset of older adults meeting criteria for frailty.The response signature was dysregulated in older adults, with the plasma cell signature induced at day 2, and was never induced in frail subjects (who were all non-responders).These results represent the first genome-wide transcriptional profiling analysis of age-associated dynamics following influenza vaccination, and implicate changes in mitochondrial biogenesis and function as a critical factor in human vaccine responsiveness.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Yale School of Medicine, New Haven, CT 06520, USA.

ABSTRACT
To elucidate gene expression pathways underlying age-associated impairment in influenza vaccine response, we screened young (age 21-30) and older (age≥65) adults receiving influenza vaccine in two consecutive seasons and identified those with strong or absent response to vaccine, including a subset of older adults meeting criteria for frailty. PBMCs obtained prior to vaccination (Day 0) and at day 2 or 4, day 7 and day 28 post-vaccine were subjected to gene expression microarray analysis. We defined a response signature and also detected induction of a type I interferon response at day 2 and a plasma cell signature at day 7 post-vaccine in young responders. The response signature was dysregulated in older adults, with the plasma cell signature induced at day 2, and was never induced in frail subjects (who were all non-responders). We also identified a mitochondrial signature in young vaccine responders containing genes mediating mitochondrial biogenesis and oxidative phosphorylation that was consistent in two different vaccine seasons and verified by analyses of mitochondrial content and protein expression. These results represent the first genome-wide transcriptional profiling analysis of age-associated dynamics following influenza vaccination, and implicate changes in mitochondrial biogenesis and function as a critical factor in human vaccine responsiveness.

Show MeSH
Related in: MedlinePlus