From Longevity Wiki

BPIFB4 (Bactericidal/permeability-increasing fold-containing family B member 4), formerly known as LPLUNC4, is a secreted protein that in humans is encoded by the BPIFB4 gene located on chromosome 20.[1] The BPIFB4 consists of 614 amino acids and has a molecular weight of 65,055 Da. It belongs to the BPI (bactericidal/permeability-increasing protein) fold-containing superfamily, which are defined as proteins that contain domains of the BPI fold and are known as antibacterial components participating in host protection through antimicrobial, surfactant, and immunomodulatory properties.[2][1] Domains of the BPI fold act as pockets capable of binding lipids, especially phosphatidylcholine.[3]

BPIFB4 was originally found to be located in the salivary glands and later showed to play an important pathophysiological role at the system level. It is now known to also be expressed in the olfactory epithelium, mononuclear cells, germline, stem cells, progenitor cells and fetal cells.

LAV — longevity-associated variant of BPIFB4


Genome-wide association search (GWAS) of long-living individuals has shown that certain BPIFB4 variants are associated with exceptional longevity, in particular a variant containing four missense mutations (Val229/Thr281/Phe488/Thr494-BPIFB4). This nucleotide polymorphism has an allele frequency in the European population of 29.5% called LAV (longevity-associated variant), wherein LAV homozygous genotype reaches 14% in centenarians.[4][5] The most common variant (Ile229/Asn281/Leu488/Ile494-BPIFB4) called wild-type-WT-BPIFB4, is found in 66% of population.

Cellular localization and activation of LAV-BPIFB4

WT-BPIFB4 and LAV-BPIFB4 have different subcellular localization. In particular WT-BPIFB4 shows nuclear localization, while LAV-BPIFB4 is mostly cytoplasmic.[4] Cytoplasmic localization of LAV-BPIFB4 allows its activation by phosphorylation at Ser75 of BPIFB4, and can be bound by 14-3-3 proteins (which can also bind the longevity FOXO proteins). 14-3-3 proteins are capable of modulating signalling pathways related to transcriptional changes, apoptosis, autophagy, endoplasmic reticulum stress, cell cycle regulation, mitochondrial function and excitation–contraction coupling in smooth muscle.[6]

The BPIFB4/14-3-3 complex can recruit the longevity-associated heat-shock protein 90 kDa (Hsp90), known to regulate proteostasis.[7] This induces eNOS phosphorylation in Ser1177, resulting in NO (nitric-oxide) production in endothelial cells.[7][8] Moreover, LAV-BPIFB4 is phosphorylated by protein kinase C alpha (PKCα) in Ser75, which activates calcium mobilization, and in turn determines PKCα activation, generating a feed-forward mechanism. [7]

LAV-BPIFB4 in longevity

Healthily aged long-living individuals have significantly higher serum BPIFB4 than do frail individuals.[9] BPIFB4 mRNA levels are also elevated in the total circulating mononuclear cells fraction from healthy long-living individuals, as compared with non-healthy frail individuals.[10]

Elderly people have a reduced exercise tolerance and a decreased left ventricle inotropic and perfusion reserve.[11] In addition, adrenergic responsiveness is altered with ageing. This may partly be explained by the fact that older failing hearts have a deficit in BPIFB4, which is associated with the scarcity of capillaries and surrounding pericytes.[12]

In vivo studies demonstrated the delivery of the LAV-BPIFB4 gene through an adeno-associated virus (AAV) carrying a liver-specific promoter exerted broad protection in rodent models of cardiovascular disease.[12] This gene transfer method allows sustained expression of secreted therapeutic proteins in the liver and systemic circulation for cross-correction of disease in other body districts.[12] In a study on elderly mice of both sexes LAV-BPIFB4 treatment reduced myocardial fibrosis and rescued cardiac function and myocardial perfusion by improving microvasculature density and pericyte coverage.[12] Moreover, gene therapy by systemic adeno-associated viral vector-mediated LAV-BPIFB4 gene transfer counteracted the development of vascular atherosclerosis in ApoE knockout mice fed with a high fat diet.[13] This might make LAV-BPIFB4 a suitable candidate tool for the treatment of atherosclerosis and its related CVD complications. Furthermore, LAV-BPIFB4 gene therapy caused a temporary reduction in systolic blood pressure in diabetic and non-diabetic aged mice, with pressure values returning to baseline by day 6 until the end of the study, albeit the benefit on endothelial-mediated vasorelaxation persisted up to a month after treatment.[14] In old mice, LAV-BPIFB4 gene transfer delayed frailty progression.[9] Gene therapy with LAV-BPIFB4 prevented cardiac deterioration in middle-aged (18-month-old) mice and rescued cardiac function and myocardial perfusion in older mice by improving microvasculature density and restoration of pericyte function and pericyte-endothelial cell interactions. [12] "Translated to the human condition, this recovery of contractility indexes seen in older mice would correspond to rewinding the heart’s biological clock by more than 10 years."[15]

Molecular mechanism of LAV-BPIFB4

Therapeutic effects of LAV-BPIFB4 may be due to its regulation of calcium via CXCL12 chemokine receptor CXCR4-SDF1 pathway.[13][14] LAV-BPIFB4 exerted an anti-inflammatory and pro-resolving macrophage M2-polarizing effect via a CXCR4-dependent mechanism and a reduction in T-cell activation.[13][16]

The mechanism by which LAV-BPIFB4 gene therapy supplementation restores pericyte function and pericyte-endothelial cell interactions requires the involvement of a nucleolar protein called nucleolin.[12]

LAV-BPIFB4 activity might also be preventing the aging-specific disruption of NAD+ homeostasis.[17] This seems to occur via the reduction of upbuilding CD38+ inflammatory cells, which generally accumulate in tissues during chronological aging and actively destroy NAD+.[17] Long-living-individuals LAV-carriers, were characterized by having high NAD+ levels.[17]

RV — aging-associated variant of BPIFB4

A Rare Variant (Ile229/Asn281/Phe488/Thr494-BPIFB4) known as RV-BPIFB4, is found in up to 4% of the population.[4] RV-BPIFB4 might server as a novel marker of vascular dysfunction and hypertension, and potentially of accelerated aging.

Human carriers of RV-BPIFB4 have significantly increased diastolic blood pressure, and exposure of ex-vivo human vessels to a human recombinant RV-BPIFB4 protein has shown to negatively modulate endothelial function and eNOS activation.[18] Endothelial nitric oxide synthase (eNOS) is a key vasodilating enzyme regulating organismal homeostasis and associated to exceptional longevity.[19] When eNOS activity is reduced, it leads to vascular disease.[20] Furthermore, forced expression of RV-BPIFB4 in WT mice by dint of injected RV-BPIFB4-encoding AAV vector, evoked vascular eNOS dysfunction and increased basal systolic blood pressure to a degree similar to that seen in eNOS-deficient mice.[18]

Therefore, RV-BPIFB4 leading to hypertension and reduced eNOS, both markers of shorter remaining lifespan and increased aging, is another indicator of BPIFB4 variants being associated to longevity.


  1. 1.0 1.1 Bingle, C. D., & Gorr, S. U. (2004). Host defense in oral and airway epithelia: chromosome 20 contributes a new protein family. The international journal of biochemistry & cell biology, 36(11), 2144-2152. PMID: 15313462 DOI: 10.1016/j.biocel.2004.05.002
  2. Bingle, C. D., Seal, R. L., & Craven, C. J. (2011). Systematic nomenclature for the PLUNC/PSP/BSP30/SMGB proteins as a subfamily of the BPI fold-containing superfamily. PMC3196848 DOI: 10.1042/BST0390977
  3. Bülow, S., Zeller, L., Werner, M., Toelge, M., Holzinger, J., Entzian, C., ... & Gessner, A. (2018). Bactericidal/permeability-increasing protein is an enhancer of bacterial lipoprotein recognition. Frontiers in immunology, 9, 2768. PMID: 30581431 PMC6293271 DOI: 10.3389/fimmu.2018.02768
  4. 4.0 4.1 4.2 Villa, F., Carrizzo, A., Spinelli, C. C., Ferrario, A., Malovini, A., Maciąg, A., ... & Puca, A. A. (2015). Genetic analysis reveals a longevity-associated protein modulating endothelial function and angiogenesis. Circulation research, 117(4), 333-345. Circ Res; 117(4): 333–345. PMC: 5496930 doi :10.1161/CIRCRESAHA.117.305875
  5. Spinelli, C. C. (2014). La Variante Associata alla Longevità (LAV) del gene BPIFB4 modula eNOS e la funzionalità vascolare. TESI DI DOTTORATO DI RICERCA. TUTORE: Prof. S. BOSARI; CO-TUTORE: Dott. A. PUCA.
  6. Thompson, W. C., & Goldspink, P. H. (2022). 14–3-3 protein regulation of excitation–contraction coupling. Pflügers Archiv-European Journal of Physiology, 474(3): 267–279. PMID: 34820713 PMC8837530 DOI: 10.1007/s00424-021-02635-x
  7. 7.0 7.1 7.2 Spinelli, C. C., Carrizzo, A., Ferrario, A., Villa, F., Damato, A., Ambrosio, M., ... & Vecchione, C. (2017). LAV-BPIFB4 isoform modulates eNOS signalling through Ca2+/PKC-alpha-dependent mechanism. Cardiovascular Research, 113(7), 795-804. PMID: 28419216 PMC5437365 DOI:10.1093/cvr/cvx072
  8. Kraehling, J. R., & Sessa, W. C. (2015). Enhanced eNOS activation as the fountain of youth for vascular disease: is BPIFB4 what ponce de leon was looking for?. Circulation research, 117(4), 309-310. PMID: 26227874 PMC: PMC4676071 DOI: 10.1161/CIRCRESAHA.115.307020
  9. 9.0 9.1 Malavolta, M., Dato, S., Villa, F., De Rango, F., Iannone, F., Ferrario, A., ... & Puca, A. A. (2019). LAV-BPIFB4 associates with reduced frailty in humans and its transfer prevents frailty progression in old mice. Aging (Albany NY), 11(16), 6555. PMID: 31461407 PMC6738439 DOI:10.18632/aging.102209
  10. Spinetti, G., Sangalli, E., Specchia, C., Villa, F., Spinelli, C., Pipolo, R., ... & Puca, A. A. (2017). The expression of the BPIFB4 and CXCR4 associates with sustained health in long-living individuals from Cilento-Italy. Aging (Albany NY), 9(2), 370. PMID: 28121621 PMC5361669 DOI:10.18632/aging.10115
  11. Kokkinos, P., Faselis, C., Samuel, I. B. H., Pittaras, A., Doumas, M., Murphy, R., ... & Myers, J. (2022). Cardiorespiratory fitness and mortality risk across the spectra of age, race, and sex. Journal of the American College of Cardiology, 80(6), 598-609. PMID: 35926933 DOI: 10.1016/j.jacc.2022.05.031
  12. 12.0 12.1 12.2 12.3 12.4 12.5 Cattaneo, M., Beltrami, A. P., Thomas, A. C., Spinetti, G., Alvino, V., Avolio, E., ... & Madeddu, P. (2023). The longevity-associated BPIFB4 gene supports cardiac function and vascularization in aging cardiomyopathy. Cardiovascular Research. PMID: 36635236 DOI:10.1093/cvr/cvad008
  13. 13.0 13.1 13.2 Puca, A. A., Carrizzo, A., Spinelli, C., Damato, A., Ambrosio, M., Villa, F., ... & Vecchione, C. (2020). Single systemic transfer of a human gene associated with exceptional longevity halts the progression of atherosclerosis and inflammation in ApoE knockout mice through a CXCR4-mediated mechanism. European Heart Journal, 41(26), 2487-2497. PMID: 31289820 PMC7340354 DOI: 10.1093/eurheartj/ehz459
  14. 14.0 14.1 Dang, Z., Avolio, E., Thomas, A. C., Faulkner, A., Beltrami, A. P., Cervellin, C., ... & Madeddu, P. (2020). Transfer of a human gene variant associated with exceptional longevity improves cardiac function in obese type 2 diabetic mice through induction of the SDF‐1/CXCR4 signalling pathway. European journal of heart failure, 22(9), 1568-1581. PMC8220375 doi:10.1002/ejhf.1840
  15. Haridy R. (2023). Unique gene mutation in superagers could rewind heart age by a decade. New Atlas.Medical. January 23, 2023
  16. Di Pardo, A., Ciaglia, E., Cattaneo, M., Maciag, A., Montella, F., Lopardo, V., ... & Puca, A. A. (2020). The longevity-associated variant of BPIFB4 improves a CXCR4-mediated striatum–microglia crosstalk preventing disease progression in a mouse model of Huntington’s disease. Cell death & disease, 11(7), 1-16. PMID: 32683420 PMC7368858 DOI:10.1038/s41419-020-02754-w
  17. 17.0 17.1 17.2 Ciaglia, E., Lopardo, V., Montella, F., Carrizzo, A., Di Pietro, P., Malavolta, M., ... & Puca, A. A. (2022). Transfer of the longevity-associated variant of BPIFB4 gene rejuvenates immune system and vasculature by a reduction of CD38+ macrophages and NAD+ decline. Cell death & disease, 13(1), Article number: 86. PMID: 35087020 PMC8792139 DOI:10.1038/s41419-022-04535-z
  18. 18.0 18.1 Vecchione, C., Villa, F., Carrizzo, A., Spinelli, C. C., Damato, A., Ambrosio, M., ... & Puca, A. A. (2017). A rare genetic variant of BPIFB4 predisposes to high blood pressure via impairment of nitric oxide signaling. Scientific reports, 7(1), PMC5574984 DOI:10.1038/s41598-017-10341-x
  19. Puca, A. A., Carrizzo, A., Ferrario, A., Villa, F. & Vecchione, C. Endothelial nitric oxide synthase, vascular integrity and human exceptional longevity. Immunity & ageing: I & A. 9, 26 (2012).
  20. Forstermann, U. & Munzel, T. Endothelial nitric oxide synthase in vascular disease: from marvel to menace. Circulation. 113, 1708–14 (2006).

For further information

  • Montella, F., Lopardo, V., Cattaneo, M., Carrizzo, A., Vecchione, C., Puca, A. A., & Ciaglia, E. (2022). Role of BPIFB4 in Immune System and Cardiovascular Disease: The Lesson from Centenarians. Transl. Med. UniSa, 24(1), Article 2. https://doi.org/10.37825/2239-9747.1024
  • Lopardo, V., Conti, V., Montella, F., Iannaccone, T., Esposito, R. M., Sellitto, C., ... & Ciaglia, E. (2022). Gender Differences Associated with the Prognostic Value of BPIFB4 in COVID-19 Patients: A Single-Center Preliminary Study. Journal of personalized medicine, 12(7), 1058. https://doi.org/10.3390/jpm12071058
  • Ciaglia, E., Lopardo, V., Montella, F., Sellitto, C., Manzo, V., De Bellis, E., ... & Puca, A. A. (2021). BPIFB4 circulating levels and its prognostic relevance in COVID-19. The Journals of Gerontology: Series A, 76(10), 1775-1783.
  • Puca, A. A., Lopardo, V., Montella, F., Di Pietro, P., Cesselli, D., Rolle, I. G., ... & Ciaglia, E. (2022). The Longevity-Associated Variant of BPIFB4 Reduces Senescence in Glioma Cells and in Patients’ Lymphocytes Favoring Chemotherapy Efficacy. Cells, 11(2), 294. https://doi.org/10.3390/cells11020294
  • Cattaneo, M., Aleksova, A., Malovini, A., Avolio, E., Thomas, A., Alvino, V. V., ... & Puca, A. A. (2023). BPIFB4 and its longevity-associated haplotype protect from cardiac ischemia in humans and mice. Cell Death & Disease, 14(8), 523. PMID: 37582912 PMC10427721 DOI: 10.1038/s41419-023-06011-8