key: cord-1017348-yuk8i9ke authors: Palmieri, Beniamino; Vadalà, Maria; Roncati, Luca; Garelli, Andrea; Scandone, Francesco; Bondi, Moreno; Cermelli, Claudio title: The Long‐Standing History of Corynebacterium Parvum, Immunity and Viruses date: 2020-05-30 journal: J Med Virol DOI: 10.1002/jmv.26100 sha: 5dec374e5c53ea616bb485c048d1b420400a67a3 doc_id: 1017348 cord_uid: yuk8i9ke We report a review of all the experimental and clinical studies performed in the last 60 years on the antiviral activity of inactivated Corynebacterium Parvum (Cutibacterium acnes). This bacterium has been originally investigated and used for its oncolytic properties linked to immunomodulating activity, but the interest to successfully prevent and treat bacterial, fungal and viral infections and lethality, uprising the innate immunity barriers produced many experimental models and very few clinical studies. The dramatic defenceless situation due to impending CoViD‐19 pandemic claims to exhume and highlight this aspecific strategy in preventive and therapeutic settings; as a matter of fact no new or mutated virus can potentially escape to this strong innate immune surveillance strengthened by adequate C.parvum protocols. This article is protected by copyright. All rights reserved. Cutibacterium acnes (formerly known as Propionibacterium acnes or Corynebacterium parvum) has been widely investigated as far as the skin microbiota environment [1]. In 1900, it was identified as Bacillus acnes [2] ; in the following years, because of its clubshaped appearance it was classified as Corynebacterium but, its mainly anaerobic metabolism and biochemical characteristics were more similar to propionic acid bacteria [3] . For this reason, the name Propionibacterium acnes (P. acnes) was successfully maintained up to 2016, when it was reclassified as Cutibacterium acnes due to peculiar genomic adaptive changes [4] . Propionibacterium acnes is a Gram-positive anaerobic bacillus that belongs to the normal cutaneous microbiota provided of immunomodulatory activity when used as a heat-or phenol-killed suspension [5] . Among its main biological activities, P. acnes promotes macrophage activation [6, 7] , displays oncolytic properties [8] [9] [10] and is an effective adjuvant when added to normal vaccines enhancing soluble and cell mediated immune response [11, 12] ; all these properties boost up the infection resistance observed after intraperitoneal or subcutaneous administration [13] [14] [15] [16] . The mechanisms responsible for the modulating effects of P. acnes on both innate and acquired immunity are mediated by interferon and proinflammatory cytokines, and are related on Toll-like receptor 2 (TLR2), Toll-like receptor 9 (TLR9), and Myeloid differentiation primary response 88 (MyD88) receptors [17] [18] [19] ; they also enhance the Th1 population function [20, 21] . The experimental in vitro/in vivo and clinical studies on immunomodulating properties of C.parvum go back to 1964 when Halpern first discovered and published his experience of This article is protected by copyright. All rights reserved. reticuloendothelial stimulation by Corynebacterium parvum injection, with enlargement of spleen and liver that reverted after the peak at 14 day, and whose reticuloendothelial hyperplasia cleared quickly intravenous delivered carbon particles [6] . In 1966 the same author and coworkers demonstrated that mice pre-treated with C.parvum were refractory to priming of tumour cells transplant [8] . In 1977 Geniteau et al published a paper, entitled: Effect of Corynebacterium parvum on various viral infections in the mouse [22] . In the same year, Glasgow and coworkers injected C.parvum in mice 7 to 10 days before inoculation of Herpes Simplex Virus hominis type 2 (HSV-2), Murine Semliki Forest Virus (SFV) [23] . The different groups of animals were highly protected against lethal infections. In the encephalomyocarditis experiment, the virus infection was counteracted either by the intraperitoneal, or the respiratory route of C. acnes administration, with a systemic, rather than local effect. C. acnes was not effective in new borne animals, probably requiring maturation of the soluble and cell mediated immune responses. The lymphoreticular system played a central role as demonstrated by transfer of enhanced resistance against HSV-2 to recipient animals infused with peritoneal exudate cells harvested from C. acnespretreated mice. The same cells inhibited in vitro the herpetic virus colonization into target cells. In this experimental study the activated macrophages were the key of immune modulating resistance to viral infections [23] . Also, Kirchner et al (1978) reported protective effect of C.parvum in normal and immunosuppressed mice [24] . In this same year, Szmigielski et al demonstrated that the intraperitoneal injection of C.parvum in HSV-2 encephalomyelitis reduced the mortality rate from 90 to 30% [25] . The same author neutralized the mortality of varicella and hepatitis B Virus (HBV) infection in proper experimental models [26] . Papaevangelou et al [27] investigated the immune potentiating effect of the intradermal administration of four doses (0.25 ml) of a standard suspension of killed C. parvum (2 This article is protected by copyright. All rights reserved. Accepted Article mg/ml) in 10 asymptomatic chronic Hepatitis B surface antigen (HBsAg) carriers, compared with 11 persons with antibodies to HBsAg (anti-HBs) and six without HBsA g or anti-HBs. HBsAg, anti-HB s, and leukocyte migration inhibition (LMI) studies were performed in pre-and post-inoculation blood samples. C. parvum produced a substantial increase of anti-HB s titre in persons with pre-existing anti-HB immunity. However, anti-HBs responses were not induced in carriers. HBsAg was not eliminated and its titre remained practically unchanged in chronic carriers. The conclusive hypothesis is, that the specific defect in the immune response to HBsAg in carriers ought to be at the B cell level. Kobus and Szmigielski demonstrated that subcutaneous C.parvum injection simultaneously with live attenuated viral vaccines achieved an higher immunity rate and protection in several experiments [28] . Teixeira and coworkers (2018) evaluated the adjuvant role of C.Parvum in BALB/c mice with HIVBr18, a DNA vaccine containing 18 CD4 + T cell epitopes from Human Immunodeficiency Virus (HIV), specifically and steadily uprising the CD4 + and CD8 + T cell responses [29] . The C.parvum added to vaccine administration increased the proliferation of HIV-1-specific CD4 + and CD8 + T lymphocytes, the polyfunctional profile of CD4 + T cells, the production of interferon gamma (IFN-γ), and the number of recognized vaccine-encoded peptides; the immunological reinforcement was more striking using the whole dead C.parvum as adjuvant compared with its bacterial polysaccharides extract; IFN-γ production as well as T cells proliferation were maintained up to ten weeks after the challenge with the whole dead bacterium [29] . Since it has been successfully used as an adjuvant in experimental trials, P.acnes can be a suitable candidate for integration in human vaccines to enhance the immune response [30, 31] . This article is protected by copyright. All rights reserved. Schindler and coworkers observed a protective effect when C. parvum was given 2h before or 2h after viral infection with Mouse Hepatitis Virus (MHV) a Coronavirus strain 3 (MHV-3) [35] . He suggested also that C. parvum determined a significant decrease of virus yield in cultures of peritoneal exudate cells infected with MHV3. He showed that contemporary intraperitoneal injection of the virus and of C.parvum prevented the infection with activation of interferon and natural killer cells but also with a direct action upon macrophages who are the primary target of virus replication. In fact Belyavsky et al (1998) demonstrated that Coronavirus MHV-3 produces fulminant lethal infection in fully susceptible BALB/c mice with massive-apoptosis in macrophages that play a key role in the virus induced liver failure [36] . Mak and coworkers (1983) showed that intranasal administration of 350 micrograms C.parvum 3 days in advance protected different mice strains (c57BL/6j, BALB/CBalb/cnu+NU+) from influenza lethality [37] . A substantial reduction of the intrapulmonary copies of virus was detected in parallel to the level of interferon and macrophage activation in the lung. Despite the increase in macrophage content, the level of specific immune responses to infection, such as cytotoxic T-cell activity, delayed-type hypersensitivity (DTH) reaction, and anti-hemagglutinin antibody, remained unchanged by C. parvum treatment so that the major if not exclusive effect of this treatment was supposed to be an uplift in the general components of the immune system. Zgórniak-Nowosielska et al (1989) achieved strong resistance against Vaccinia Viruses (VACV) and Herpes Virus-1 with C.parvum pretreatment [38] . Splenomegaly was induced by the procedure, but the specific antibodies titer was unchanged suggesting enhancement of natural immunity rather than specific soluble immunity stimulation. Kounoue et al (1987) prevented the virus induced encephalomyocarditis of mice with a 3-14 tailored pretreatments of C.parvum; this therapy prevented pancreas colonization by virus, causing insulae destruction and diabetes [39] . Cohen and coworkers (1984) treated Ectromelia Virus (ECTV) infected mice with intraperitoneal C.parvum in comparison with mineral oil adjuvant injection [40] . The protection was obtained through macrophage uptaking and killing of the viruses and interferon induction [40] . Mayr et al Megid, one year before, had measured the natural killer (NK) activity and lethality in the same mice model exposed to rabies virus and treated with C.acnes; the mice were progressively killed and the spleen cells rescued and analysed [47] . Higher NK activity and better survival rate were observed in the P. acnes cohort outlining this lymphocyte population as the key mechanism of rabies virus neutralization. In a mouse model of vaccination, the same author (2006) This article is protected by copyright. All rights reserved. Kalis and coworkers (2005) focused primarily on the bacterium-host immune cell interaction in order to better understand the immunomodulating mechanisms of killed C. parvum preparation through systemic and organ specific (lung) innate immunity: the receptor type involved in this interaction is Toll-like receptor 9 (TLR9) [18] . [56] . They express only TLR7, 8 and 9, but not TLR2, TLR3, TLR4 and TLR5. Therefore, they are supposed through TLR9 and TLR7 to play a primary role in DNA and RNA virus to some DNA and RNA viruses, preventing the infections either shaping and upgrading innate immunity, or translating the innate response to adoptive immunity [56] . pDCs produce massive amounts of type I interferon, activating NK cells, NKT cells, B cells, T cells, and myeloid uncommitted DCs and then complete the suppression and clearance of the virus particles [57] . C.Parvum probably recruits and activates these cells with chemotactic signal, enhancing the defence against viruses. We suppose that in humans, as in the experimental animal models, the dead bacterium acts initially as a strong TLR9 agonist, activating macrophages and promoting high levels of Th1, TNF, interferon, and natural killer cells to clear the infecting pathogens. Another intriguing interplay between viruses and C.parvum is the pivotal role of vimentin. We know from dermatological investigations that inflammation of sebaceous glands and keratinocytes by Cutibacterium acnes involves a complex cytokines cascade, and that live C. acnes enters into monocytes and tissue cells and can survive and proliferate interacting with vimentin network, a component of the cytoskeleton [58, 59] . P. acnes invades the prostate cells that express vimentin, which is able to activate immune response against the bacterium through the Nuclear factor-κB (NFkB) intracellular innate immune signalling [60] . PRRs, once alerted, activate the inflammasome and NF-kB cascade to destroy the hosts [68] . Pathogenic viruses, however, can try to overwhelm these defence mechanisms, inactivating the vimentin-PRRs complex by post-translational modifications. In this perspective, the puzzling competition between the subcutaneous injected dead C. parvum and virus infections plays a determinant role to balance the innate immune defence mechanisms and virulence factors. In conclusion, the long experimental history documenting a wide spectrum antiviral activity of C. parvum lysates, both in vitro and in vivo, besides a limited number of clinical trials, strongly supports the hypothesis that also SARS-CoV-2 virus can be effectively neutralized by this treatment, suggesting that further investigation is worth to be carried out, mainly by clinical trials. This potential aspecific anti SARS-CoV-2 activity could efficiently prevent and treat this dramatically world spreading viral disease and help the recovery of social and productive life. Author contribution statement: Beniamino Palmieri: Conceptualization (lead); writing-original draft (lead); formal analysis (lead); writing -review and editing (equal). Propionibacterium (Cutibacterium) acnes Bacteriophage Therapy in Acne: Current Evidence and Future Perspectives. Dermatology and therapy A bacteriological and microscopical study of over 300 vesicular and pustular lesions of the skin, with a research upon the etiology of acne vulgaris The Taxonomic Position of Corynebacterium acnes Recent advances in understanding Propionibacterium acnes ( Cutibacterium acnes) in acne. F1000Research Age-related changes in the resident bacterial flora of the human face STIMULATION OF THE PHAGOCYTIC ACTIVITY OF THE RETICULOENDOTHELIAL SYSTEM BY CORYNEBACTERIUM PARVUM Tumour growth, phagocytic activity and antibody response in Corynebacterium parvum-treated mice Inhibition of tumour growth by administration of killed corynebacterium parvum Modulation of tumoricidal activity, induced in bone-marrow-derived mononuclear phagocytes by interferon gamma or Corynebacterium parvum, by interferon beta, tumor necrosis factor, prostaglandin E2, and transforming growth factor beta Effect of intracerebrally injected Corynebacterium parvum on the development and growth of metastatic brain tumor in mice Effect of Corynebacterium parvum on the class and subclass of antibody produced in the response of different strains of mice to sheep erythrocytes Adjuvant effect of the Propionibacterium acnes and its purified soluble polysaccharide on the immunization with plasmidial DNA containing a Trypanosoma cruzi gene Increased nonspecific resstance to malaria produced by administration of killed Corynebacterium parvum The effect of in vivo modulation of macrophage activities on Mycobacterium lepraemurium infection The use of non-specific immunopotentiators in experimental Trypanosoma cruzi infection Sorveglianza immunologica e immunoterapia aspecifica nel malato neoplastico. Recenti progressi in medicina, il pensiero scientifico editore Roma Activation of toll-like receptor 2 in acne triggers inflammatory cytokine responses Requirement for TLR9 in the immunomodulatory activity of Propionibacterium acnes Propionibacterium acnes induces an adjuvant effect in B-1 cells and affects their phagocyte differentiation via a TLR2-mediated mechanism CD8-dendritic cell activation status plays an integral role in influencing Th2 response development Cutting edge: dendritic cells copulsed with microbial and helminth antigens undergo modified maturation, segregate the antigens to distinct intracellular compartments, and concurrently induce microbe-specific Th1 and helminthspecific Th2 responses Immunomodulation of host resistance to experimental viral infections in mice: effects of Corynebacterium acnes, Corynebacterium parvum, and Bacille calmette-guerin Protection of mice against viral infection by Corynebacterium parvum and Bordetella pertussis Reaction of cell-mediated immunity tolocal hyperthermia of tumors. Cancers therapy by hyperthermia and radiation Protection and therapy of mice with acute and chronic experimental virus infections with propionibacterium granulosum kp-45 The effect of intradermal administration of corynebacterium parvum on the immune response to hepatitis Bs antigen Protection and therapy of experimental herpesvirus infections in mice with immunomodulating Propionibacterium avidum KP-40 and/or acyclovir Propionibacterium acnes Enhances the Immunogenicity of HIVBr18 Human Immunodeficiency Virus-1 Vaccine Adjuvant therapy of malignant melanoma Alternatives to chemotherapy and radiotherapy as adjuvant treatment for lung cancer. Lung Cancer Enhanced resistance against Junin virus infection induced by Corynebacterium parvum Antiviral properties of five strains of Corynebacterium Hebd Seances Acad Sci D Protection of mice against mouse hepatitis virus by Corynebacterium parvum. Infection and immunity Coronavirus MHV-3-induced apoptosis in macrophages Protection of mice against influenza virus infection: enhancement of nonspecific cellular responses by Corynebacterium parvum Protection of mice against vaccinia and herpes simplex virus infection by Propionibacterium acnes Encephalomyocarditis (EMC) virus-induced diabetes mellitus prevented by Corynebacterium parvum in mice Abortive ectromelia virus infection in peritoneal macrophages activated by Corynebacterium parvum Treatment of common warts with the immune stimulant Propionium bacterium parvum Propionibacterium Acnes: A Putative Immunemodulating Weapon Against the Coronavirus Impending Epidemy Plasmacytoid dendritic cells act as the most competent cell type in linking antiviral innate and adaptive immune responses CpG ODN enhance antigen-specific NKT cell activation via plasmacytoid dendritic cells The role of inflammation in the pathology of acne Interaction of Cutibacterium ( formerly Propionibacterium) acnes with bone cells: a step toward understanding bone and joint infection development. Scientific reports Propionibacterium acnes host cell tropism contributes to vimentin-mediated invasion and induction of inflammation Cell surface vimentin is an attachment receptor for enterovirus 71 Surface vimentin is critical for the cell entry of SARS-CoV The origin, transmission and clinical therapies on coronavirus disease 2019 (COVID-19) outbreak -an update on the status The intermediate filament protein, vimentin, is a regulator of NOD2 activity Vimentin regulates activation of the NLRP3 inflammasome Vimentin is an endogenous ligand for the pattern recognition receptor Dectin-1 Astrogliosis involves activation of retinoic acidinducible gene-like signaling in the innate immune response after spinal cord injury The cytoskeleton in cell-autonomous immunity: structural determinants of host defence [55 ]