Adsorption, penetration and uncoating are the first stage of viral replication. Viruses encode multiple proteins to manipulate the plasma membrane Ca²⁺ channels [VGCCs and two-pore channels (TPCs)] involved in regulating cellular Ca²⁺ uptake to increase intracellular Ca²⁺, thereby facilitating viral entry and replication. For example, human immunodeficiency virus type 1 (HIV-1) gp120 and Tat protein, and porcine rotavirus (RV), activate VGCCs to increase the levels of intracellular Ca²⁺ (Holden et al., 1999; Pérez et al., 1999; Mayne et al., 2000). The glycoprotein hemagglutinin (HA) of Alphainfluenzavirus influenzae (IAV) binds to voltage-dependent Ca²⁺ channel Ca_v1.2, triggering intracellular Ca²⁺ increase, and activates endocytosis to gain entry into cells (Fujioka et al., 2013, 2018; Figure 2A). Human alphaherpesvirus 1 (HSV)-1 infection results in a significant decrease in protein expression of Ca_v3.2 T-type Ca²⁺ channel subunit to escape detection by host cells (Zhang et al., 2019). Some VGCC blockers inhibit viral replication by inhibiting intracellular Ca²⁺ increase, such as Human gammaherpesvirus 4 (EBV), Human betaherpesvirus 5 (CMV), and flaviviruses [such as Japanese encephalitis virus (JEV), Zika virus (ZIKV), dengue virus (DENV) and West Nile virus (WNV)] (Nokta et al., 1987; Dugas et al., 1988; Wang et al., 2017). Intracellular Ca²⁺ oscillations are the trigger for viral penetration or uncoating (Baravalle et al., 2004; Danta, 2020). For porcine rotavirus, the critical step for uncoating and membrane permeabilization is a decrease in Ca²⁺ concentration of cytosol accompanying dissociation of viral Ca²⁺-stabilized proteins (Chemello et al., 2002; Salgado et al., 2018). The structure of some viruses contains a flexible surface loop that binds divalent Ca²⁺, and have been found to be essential for virus infectivity (Simpson et al., 2000).

In the 1990s, the role of the endolysosomal system for calcium storage was discovered (Genazzani and Galione, 1996). The calcium exchange and Ca²⁺-mediated functional coupling also exists at the lysosome-ER interface (Raffaello et al., 2016). The mucolipin family of transient receptor potential (TRPML) channels and TPCs are involved in the release of Ca²⁺ from lysosomes (Patel et al., 2011; Figure 1). Many viruses mobilize Ca²⁺ from the endolysosomal stores through TPCs, and some viruses are dependent on the activity of TPCs to enable capsid entry into cells using endolysosomal networks, such as Alphapolyomavirus quintihominis (MCPyV), Betapolyomavirus macacae (SV40), Middle East respiratory syndrome coronavirus (MERS-CoV) and Zaire Ebola virus (EBOV; Sakurai et al., 2015; Gunaratne et al., 2018; Dobson et al., 2020; Figure 2B).

Viral biosynthesis includes mRNA transcription, and protein and DNA or RNA synthesis and metabolism, in which Ca²⁺-signaling networks play an important role. Some viruses or viral proteins are involved in increasing cytoplasmic Ca²⁺, leading to activation of the Ca²⁺-sensitive nuclear factor of activated T cells (NFAT) to support viral RNA transcription and establish persistent infection (Figure 2C). For example, HIV accessory protein Nef cooperates with Vpr to trigger release of ER Ca²⁺ that leads to activation of NFAT (Kinoshita et al., 1997; Lahti et al., 2003). Primate T-lymphotropic virus 1 (HTLV)-1 regulatory and accessory genes p12^I activates NAFT (Ding et al., 2002), and upregulates expression of another Ca²⁺-sensitive transcription factor, P300 (Nair et al., 2006). Human gammaherpesvirus 8 (KHSV) K1 protein, is a transmembrane glycoprotein, increased cellular tyrosine phosphorylation and intracellular Ca²⁺ mobilization, activating NFAT and AP-1 transcription factor and producing inflammatory cytokines to promote viral dissemination (Lee et al., 2005). Furthermore, intracellular calcium triggers calcineurin-dependent signal transduction resulting in reactivation of latent KHSV infection (Zoeteweij et al., 2001).

Viruses directly activate calcium channels and pumps on some organelles to promote viral protein synthesis and modification. There are reports that glycoproteins of Paramyxoviridae, Flaviviridae and Togaviridae families fail to mature in SPCA1-deficient cells, Ca²⁺ is pumped into the Golgi by SPCA1, which triggers synthesis of functional viral glycoproteins that are essential for viral spread (Hoffmann et al., 2017; Figure 2D). Some respiratory viruses, such as Human orthopneumovirus (RSV), Measles morbillivirus (MV) and Human rhinovirus A, promote upregulation of TRP channels such as TRPV1, TRPA1 and TRPM8, and use TRP channels to create an intracellular Ca²⁺ environment conducive to their replication (Abdullah et al., 2014; Omar et al., 2017). DENV, hepacivirus C (HCV) and ZIKV or the purified viral envelope protein activate TRPV4-mediated Ca²⁺ influx that drives DDX3X nuclear translocation to promote viral replication; what’s more, TRPV4-DDX3X interaction regulates the nuclear export and translation of unspliced HIV-1genomic RNA (gRNA) and viral RNA metabolism (Doñate-Macián et al., 2018; Figure 2E).

Viral proteins can also bind directly to Ca²⁺ to promote viral biosynthesis. For example, Bluetongue virus (BTV) nonstructural phosphoprotein NS2 is a dedicated Ca²⁺-binding protein that significantly enhances NS2 phosphorylation, triggering viral inclusion body formation and facilitating viral replication and assembly (Rahman et al., 2020). High levels of cytosolic Ca²⁺ facilitates hepatitis B virus (HBV) core assembly, which is regulated by the viral multifunctional regulatory protein HBx (Choi et al., 2005). High Ca²⁺ levels promote Ca²⁺-dependent activation of proline-rich tyrosine kinase 2 and focal adhesion kinase; and induce NFAT expression, which supports HBV reverse transcription and DNA replication (Lara-Pezzi et al., 1998).

Most nonenveloped viruses release virus particles when the host cell is completely lysed, while enveloped viruses release virus particles by budding, which is accompanied by activation of Ca²⁺ channels and mobilization. SOCs mediated entry of Ca²⁺ is activated mainly by the membrane Ca²⁺ release-activated Ca²⁺ modulator 1(ORAI1) on the plasma membrane and stromal interaction molecule (STIM1) on the ER, which are stimulated by the depletion of internal Ca²⁺ stores. The budding process of enveloped viruses depends on the host Ca²⁺ signal mediated by SOCs (STIM1/ORAI1). Some viruses activate SOCs or depend on STIM1 and ORAI1 interaction to enhance cellular Ca²⁺ uptake for the budding of mature virus particles. Such viruses include DENV, EBOV, Marburgvirus (MARV) and Argentinian mammarenavirus (JUNV; Han et al., 2015; Dionicio et al., 2018; Figure 2F).

Apoptosis is considered to be a form of cell suicide, regulated by its own process under physiological or pathological conditions. The morphological manifestations of apoptosis are cell shrinkage, nuclear fragmentation, chromatin condensation, and formation of apoptotic bodies. Ca²⁺ is an important signaling molecule that regulates apoptosis during viral infection. On the one hand, viral infection induces increased extracellular Ca²⁺ influx and Ca²⁺ release from intracellular storage, and the continuously increased Ca²⁺ initiates apoptosis. On the other hand, the release of Ca²⁺ from storage disrupts the stability of intracellular structure, and many key components of the apoptotic system are activated. These two forms often coexist in virus-induced apoptosis.

Mitochondria, as the core organelles of apoptosis, are hijacked and utilized by viruses. Viruses trigger apoptosis by increasing mitochondrial Ca²⁺ uptake, enhancing mitochondrial membrane permeability and promoting release of apoptotic factors. Cytochrome (Cyt) C, as an activator of the caspase family, is required for caspase-dependent apoptosis. It is released into the cytoplasm and binds to apoptotic protease activating factor (Apaf)-1 to form apoptosomes, triggering the caspase cascade via caspase-9 activation (Figure 3). For example, both HBV and HCV are hepatotropic viruses that cause chronic liver disease and hepatocellular carcinoma (Scrima et al., 2018), and they promote Ca²⁺ uptake in mitochondria and lead to reactive oxygen species (ROS) production and apoptosis (Li et al., 2007). During HBV infection, HBx protein interacts with VDACs to trigger the release of Cyt C from the mitochondria, which triggers apoptosis. Some inhibitors of Ca²⁺ channels can down-regulate the proliferation of virus and avoid the occurrence of apoptosis of host cells (Chami et al., 2003; Xia et al., 2006). In the same way, enteroviral infections cause high mitochondrial Ca²⁺ overload, mitochondrial dysfunction, and apoptosis. When treated with the BAPTA-AM, a Ca²⁺ chelating agent, viral replication was also inhibited along with the alleviation of apoptosis (Brisac et al., 2010; Peischard et al., 2019). Besides, some viral proteins, such as IAV PB1-F2, induce permeabilization and destabilization of mitochondrial membranes via changes in Ca²⁺ homeostasis, leading to macromolecular leakage and apoptosis (Chanturiya et al., 2004). Whereas HCV NS5A protein promotes IP₃R degradation, inhibiting virus-induced apoptosis and establishing chronic infection (Kuchay et al., 2018). In this process, apoptosis appears to be a host suicide defense mechanism to prevent spreading of virus.

Another promoter of cell death is apoptosis-inducing factor (AIF), which mediates the regulation of caspase-independent apoptosis (Figure 3). AIF is a mitochondrial oxidoreductase with a molecular weight of about 62 kDa anchored to the inner mitochondrial membrane (IMM) in the vicinity of Complex I. when mitochondria are damaged and the mitochondrial permeability transition pore (MPTP) is open, activated AIF is released (Susin et al., 1996). Calpain is a calcium-dependent intracellular cysteine protease that cleaves mitochondrial AIF to promote AIF activation (Cao et al., 2007; Figure 4B). The released AIF recruits macrophage migration inhibitory factor (MIF) to the nucleus, fragmentating the DNA (Yu et al., 2002; David et al., 2009; Wang et al., 2016).

In conclusion, Ca²⁺ homeostasis imbalance, mitochondrial damage, and release of various apoptotic macromolecules are often associated with virus-induced apoptosis. From the perspective of virus, it exploits host resources by destroying intracellular Ca²⁺ homeostasis and regulates apoptosis to achieve the purpose of promoting its own replication. Similarly, apoptosis is also regarded by the host as an antiviral method, and the virus has achieved the establishment of persistent infection by regulating the activity of Ca²⁺ channels to prevent apoptosis.

Under physiological conditions, mitochondrial Ca²⁺ uptake is thought to serve as a safety buffer, maintaining cellular Ca²⁺ homeostasis in the event of a temporary intracellular Ca²⁺ overload, and promoting mitochondrial oxidative phosphorylation and ATP synthesis (Vasington and Murphy, 1962; Rizzuto et al., 1992). However, viral infection can lead to continuous Ca²⁺ accumulation and induce mitochondrial Ca²⁺ overload. Most of the mitochondrial Ca²⁺ overload comes from release from the ER, which is mainly because viral infection induces membrane contact between the ER and mitochondria (Vance, 2014). Mitochondria tightly associated with elements of ER can be isolated, and these membranes structures are frequently called mitochondria-associated membranes (MAMs) (Figure 1). IP₃Rs are highly concentrated Ca²⁺ channels in the MAMs and play a central role that turn on/off Ca²⁺ release from ER stores (Mikoshiba and Hattori, 2000). Ca²⁺ released by ER is transported to the mitochondrial intermembrane space through the Ca²⁺ channel VDACs on the outer mitochondrial membrane (OMM) (Gincel et al., 2001), and the complicated MCU complex involved in Ca²⁺ transport through the inner mitochondrial membrane (IMM), and collaborative molecules such as cytosolic chaperone Grp75 are involved in regulation (Baughman et al., 2011; Drago et al., 2011; Poston et al., 2013). The molecular components of the uniporter comprise the pore-forming subunits MCU and dominant-negative regulator MCUb, together with calcium sensors mitochondrial calcium uptake (MICU)1, MICU2, and attachment essential MCU regulator (EMRE) (Raffaello et al., 2013; Sancak et al., 2013; Kamer and Mootha, 2015; Kamer, 2018). Apoptosis is closely related to the Ca²⁺ status of the mitochondria, these spatial contacts between the ER and mitochondria, are often used to function as viral targets, which triggers apoptosis by regulating the movement of Ca²⁺ between the ER and mitochondria (Csordás et al., 2006; Raffaello et al., 2016; Panda et al., 2021). For example, HCV, Enterovirus C (PV) and Enterovirus B trigger apoptosis through enhancing IP₃R activity and promoting mitochondrial Ca²⁺ uptake by MCUs and VDACs (Gong et al., 2001; Brisac et al., 2010; Peischard et al., 2019). Mitochondrial Ca²⁺ efflux channels also play an important role in maintaining mitochondrial Ca²⁺ balance. Ca²⁺ efflux occurs through NCX and Ca²⁺/H⁺ antiporter (Khananshvili, 2013; Belosludtsev et al., 2019) (Figure 1).

Like IP₃Rs, RyRs are responsible for regulating release of ER Ca²⁺ (Figure 1). Both IP₃Rs and RyRs have similar tetramer structures and activation mechanisms, and are both activated by low Ca²⁺ levels and inhibited by high Ca²⁺ levels, so the low amount of Ca²⁺ released by ER further stimulates IP₃Rs and RyRs activity (Fan et al., 2015). For example, HSV, HBV, HTLV-1 and IAV infections all lead to increased intracellular Ca²⁺, mainly due to abnormal release of ER Ca²⁺ caused by activation of IP3Rs (Hartshorn et al., 1988; Ding et al., 2002; Cheshenko et al., 2003; Xia et al., 2006). When ER Ca²⁺ storage release is increased, the ER calcium ATPase (SERCA pump) is activated and allows rapid reuptake of cytosolic Ca²⁺ by the ER (Courjaret and Machaca, 2014) (Figure 1). HCV core protein overexpressed in Huh7 cells induces ER Ca²⁺ depletion by impairing SERCA pump function (Benali-Furet et al., 2005). Depletion of intracellular Ca²⁺ storage also stimulates interaction between ORAI1 on the plasma membrane and STIM1 on the ER to mediate extracellular Ca²⁺ entry (Figure 1). In addition, some viroporins form channels on the ER that directly or indirectly increase ER permeability, leading to uncontrolled outflow of Ca²⁺ into the cytoplasm (Feng et al., 2002; Griffin et al., 2003; Peischard et al., 2019; Strtak et al., 2019).

In general, viral infection induces the formation of contact sites between MAMs and mitochondria, and the abnormal release of ER Ca²⁺ leads to mitochondrial Ca²⁺ overload and initiation of apoptosis.

The effect phase of apoptosis involves decreased mitochondrial membrane potential, respiratory chain uncoupling, mitochondrial permeability enhancement, and mitochondrial swelling and rupture (Hunter and Haworth, 1979). MPTP is the release channel of apoptotic molecules (such as Cyt C and AIF) and the transport channel of mitochondrial Ca²⁺, and its increased permeability is the decisive change in the early stage of apoptosis (Szalai et al., 1999). When viruses induce mitochondrial Ca²⁺ overload, accumulation of ROS and mitochondrial membrane potential dissipation, the opening of MPTP also releases Ca²⁺ from the mitochondria. For example, Murid betaherpesvirus 1 (Panel et al., 2019), JEV (Huang et al., 2016), IAV protein PB1-F2 (Chanturiya et al., 2004) and KHSV protein K7 (Feng et al., 2002) disrupt cytoplasmic Ca²⁺ levels and destroy mitochondria.

MPTP is a protein complex that connects the cytoplasm, OMM and IMM, intermembrane space of mitochondria and mitochondrial matrix, and is composed of component and regulatory molecules. The components include VDAC, adenine nucleotide translocase (ANT), and mitochondrial matrix protein cyclophilin D (Cyp D). Cyp D is also considered to be the key to MPTP opening, and is blocked by cyclosporine A (Kokoszka et al., 2004; Li et al., 2004; Baines et al., 2007) (Figure 4A). In recent years, some new molecules that may be involved in the formation of MPTP have been discovered, such as phosphate carrier, aspartate–glutamate carrier, ornithine–citrulline carrier and mitochondrial complex I on the electron transport chain (Fontaine et al., 1998; Chauvin et al., 2001). Some viral proteins locate in the mitochondrial membrane and interact directly with the MPTP to induce its opening (Jacotot et al., 2000; Chami et al., 2003).

ROS are effective activators of MPTP opening, which is induced by increased oxidative stress, and regulate Ca²⁺ channels in the ER and mitochondria. At the initiation stage of oxidative stress apoptosis, the transient reversible opening of MPTP increases and ROS accumulation causes damage to cells (Ma et al., 2011). MPTP continues to open irreversibly, causing mitochondrial swelling and OMM rupture, and proapoptotic proteins in the mitochondrial intermembrane are released into the cytoplasm and activate apoptotic responses. At the late stage of apoptosis, ROS are not cleared due to disorder of the antioxidant system in the mitochondria and cytoplasm. High concentrations of ROS trigger oxidative stress, resulting in mitochondrial membrane potential dissipation, mitochondrial oxidative damage, and finally induction of apoptosis (Chen et al., 2003; Lin and Beal, 2006; Stowe and Camara, 2009). In conclusion, viral infection causes abnormal release of ER Ca²⁺, accumulation of ROS, and excess opening of MPTP, which play a synergistic role in apoptosis.

Among the recognized PRRs, TLRs are the most extensive and oldest form of pathogen recognition, and can recruit multiple adaptor proteins to activate transcription factors, including NF-κB, activating protein (AP)-1, and interferon (IFN) regulatory factor (IRF) family members, to cause a further inflammatory reaction and IFN-dependent antiviral immune response (Nie et al., 2018). TLRs and Ca²⁺ signaling interact with and regulate each other. TLRs (including TLR2, TLR3, TLR4, TLR7, TLR8 and TLR9) have been demonstrated to participate in cytokine production, activation of immunocytes, inflammation, and antiviral innate responses by regulating Ca²⁺ signaling (Tauseef et al., 2012; De Dios et al., 2020; Zhao et al., 2020). In particular, TLR4, which recognizes bacterial lipopolysaccharides (LPSs), is widely reported to alter cytoplasmic Ca²⁺ levels and activates Ca²⁺ signaling by regulating various Ca²⁺ channels. For example, endotoxin activates transient receptor potential canonical (TRPC) channels to induce Ca²⁺ entry in endothelial cells, secondary to TLR4-induced diacylglycerol generation (Feske et al., 2012). TLR3, TLR7, and TLR8 are RNA sensors that recognize immune-stimulated RNA and initiate downstream signals that increase the production of inflammatory cytokines and type I IFN (IFN-I). These processes are still accompanied by extracellular Ca²⁺ influx or release of Ca²⁺ from the ER and activation of associated Ca²⁺ channels. Especially when HIV infects CD4⁺ T cells, TLR7 induces increased cytoplasmic Ca²⁺, sensitizing activated T cell 2 nuclear factor (NFATc2) through calcination, thereby promoting HIV replication (Dominguez-Villar et al., 2015). By contrast, Ca²⁺ signaling also modulates TLR signaling by affecting expression of different TLR molecules or controlling TLR activation in different ways. High Ca²⁺ upregulates mRNA expression of TLR3 and other dsRNA sensors to augment antiviral activity in epidermal keratinocytes (Yamamura et al., 2018). Extracellular Ca²⁺ influx by STIM1-operated Ca²⁺ channels transmit the information of TLR stimuli to initiate innate immune responses (Tang et al., 2017). In general, TLRs interact with intracellular Ca²⁺ signals through different molecular mechanisms. It is important to note that our understanding of TLR-mediated Ca²⁺-signal-dependent cellular functions in different pathological processes is still insufficient. In particular, the effect of TLRs on Ca²⁺ signal regulation in viral infection needs more investigation.

RLRs are RNA sensor molecules that recognize the double-stranded RNAs (dsRNA), three of which are encoded in the human genome [RIG-I, melanoma differentiation-associated gene 5 (MDA5), and laboratory of genetics and physiology 2 (LGP2)], and enhance IFN production and play an important role in RNA virus infections (Yoneyama et al., 2005; Onomoto et al., 2021). RIG-I and MDA5 contain a tandem caspase recruitment domain, and interact with the mitochondrial antiviral signaling protein (MAVS) to activate IRF-3/7 and NF-κB, enhance IFN production, and promote transcriptional activation of proinflammatory cytokines (Yoneyama et al., 2015; Rehwinkel and Gack, 2020). Similarly, some data suggest that Ca²⁺ is a key regulator of the RLR pathway by regulating expression of many molecules involved in RLR signaling, or via regulation of Ca²⁺ channels. Studies have found that high levels of Ca²⁺ induce dsRNA sensors like MDA5 and RIG-I (Yamamura et al., 2018). In addition, the effects of some Ca²⁺ channels on RIG-I and MDA5 expression are related to Ca²⁺ signaling; for example, the Cav1.2 channel (a type of L-type Ca²⁺ channel) increases expression of RIG-I and MDA5 (Tammineni et al., 2018). Many reports have suggested that Ca²⁺ signaling plays an important role in the activation of RLR pathways during RNA virus infections. For example, IAV induces production of ROS to facilitate interaction of viral M2 protein with MAVS by increasing Ca²⁺ levels (Wang R. et al., 2019). Murine respirovirus can activate IRF3/7 by stimulating the ER to release Ca²⁺ through the RyR channel (Sermersheim et al., 2020). However, some studies have found that viruses recruited calcineurin, which inhibited TANK-binding kinase 1 (TBK1) phosphorylation and leading to reduced IFN-I production. Vesicular Stomatitis Virus (VSV) and HSV can restrict RLR-pathway-related antiviral innate immune response by targeting this mechanism (Huang et al., 2018). Pestivirus C infection can increase intracellular-Ca²⁺-level-induced autophagy through calcium/calmodulin dependent protein kinase 2 to suppress MAVS and decrease IFN-I production (Xie et al., 2021). Notably, the Ca²⁺ signaling pathway contributes to the regulation of RLR-mediated innate immunity, while there is no clear evidence that RLRs can control Ca²⁺ signaling. In addition, Ca²⁺ signaling can promote RLR-mediated innate immune response and reverse it in different viral infections. Therefore, it is important to illustrate the exact role of Ca²⁺ signaling in RNA virus regulation of RLRs in the future.

Mammalian cells recognize double-stranded DNA (dsDNA) to produce type I interferons (IFNs), suggesting that cytosolic DNA sensing is the important mechanism by which the innate immune system detects pathogens (Stetson and Medzhitov, 2006; West et al., 2015; Erdal et al., 2017). Several cytosolic DNA sensors, including cyclic GMP–AMP synthase (cGAS), melanoma 2, and DNA-dependent IFN regulatory activator, have been identified as involved in immune responses. An increasing number of studies on the activation of type I IFNs response by viral infection have focused on the cGAS–STING axis, and recent studies have shown that Ca²⁺ and related signaling proteins regulate cGAS–STING (Mathavarajah et al., 2019). cGAS detects reverse transcription of retroviral RNA, aberrant release of viral DNA during infection, and damage to host genomic DNA or mitochondrial DNA (Gao et al., 2013; Sun et al., 2013; Kanneganti et al., 2015). Recognition of cytosolic DNA by cGAS results in the production of second messenger cGAMP that binds to stimulator of interferon genes (STING) (Ishikawa and Barber, 2008). Activation of STING interacts with TBK1, and then STING functions as a scaffold protein for TBK1 and IRF3 assembly to stimulate phosphorylation of IRF3 and nuclear translocation and induces the transcription of IFN genes (Fitzgerald et al., 2003; Tanaka and Chen, 2012).

The current study has demonstrated that STING has a Ca²⁺-binding site; when STING forms a homodimer, two ions are shared between the two monomers of the protein, so Ca²⁺ directly participates in the regulation of STING activation (Shu et al., 2012). STING interacts with some Ca²⁺ channels to regulate Ca²⁺ flux during its activation; for example, STING interacts with Ca²⁺ transporters VDAC to facilitate Ca²⁺ uptake by mitochondria. In the resting state, STING is located in the ER and binds Ca²⁺-sensing transmembrane protein STIM1 and interacts with SERCA (Lee et al., 2013; Srikanth et al., 2019). In addition, changes in cytosolic Ca²⁺ caused by viral infection facilitate STING activation. It has been found that BAPTA-AM (an extracellular Ca2+ chelator)-mediated Ca²⁺ depletion and ionomycin-mediated Ca²⁺ elevation suppress STING-mediated IFN-β production. The function of STING is also restricted when virus-infected cells are treated with the IP₃R inhibitor 2-APB or SERCA inhibitors (Hare et al., 2015; Kwon et al., 2018; Mathavarajah et al., 2019). There is growing evidence that the cGAS–STING axis is one of the cellular pathways controlled by Ca²⁺ signaling, and the mechanism of this pathway regulating IFN-I response in microbial infection and viral diseases will be carefully studied in the future.

The NLR family includes a variety of specific cytoplasmic sensors that detect invasive pathogens and endogenous danger signals. NOD-like receptor protein (NLRP)3 is one of the best-identified DNA sensors associated with inflammasomes in the NLR family, and is activated by invading pathogens or endogenous danger signals, leading to the formation of NLRP3 inflammasomes. The NLRP3 inflammasome is a multiprotein platform that includes NLRP3, apoptosis-associated speck-like protein (ASC) and pro-caspase-1, which leads to caspase-1-dependent secretion of proinflammatory cytokines IL-1β and IL-18 (Wen et al., 2013; Liu et al., 2018). The role of Ca²⁺ signaling in NLRP3 inflammasome activation has been widely reported. Lots of evidence show that Ca²⁺ from the extracellular environment, ER, and lysosome promote the formation and activation of NLRP3 inflammasome (Swanson et al., 2019; Li et al., 2021). For example, the cytosolic Ca²⁺ mediated by ion channels TRPA1 and TRPV1 facilitate the activation of the NLRP3 inflammasome (Wang M. et al., 2019); The release of the Ca²⁺ from ER through RyRs and IP₃Rs are also observed to sensitize the NLRP3 inflammasome (Triantafilou et al., 2013a); Emanate from lysosomal Ca²⁺ stores regulate the production of pro-IL-1β by calcineurin, contributes to the activation of the NLRP3 inflammasome (Weber and Schilling, 2014). Until now, some data prove that viroporins enhance cytosolic Ca²⁺ by altering organelle membrane permeability, which promotes activation of NLRP3 inflammasomes. For example, Human rhinovirus B protein 2B targets ER and Golgi complex trigger Ca²⁺ release to stimulate NLRP3 inflammasome activation (Triantafilou et al., 2013b). Similarly, foot-and-mouth disease virus (FMDV) 2B protein as viroporins also promotes the flux of Ca²⁺, thereby stimulating NLRP3 inflammasome activation (Zhi et al., 2020). What’s more, severe acute respiratory syndrome coronavirus (SARS-CoV) envelope protein also activates the NLRP3 inflammasome by forming protein-lipid channels in ER/Golgi membranes to osmose Ca²⁺ (Nieto-Torres et al., 2015). So far, how intracellular Ca²⁺ stimulates NLRP3 activation is not fully understood. In the future, more investigations are needed to uncover the mechanisms by which Ca²⁺ mobilization induces NLRP3 inflammasome activation.