Mollusk Hemocyanins as Natural Immunostimulants in Biomedical Applications

Becker, María; Arancibia, Sergio; Salazar, Fabián; DelCampo, Miguel; Ioannes, Alfredo De

doi:10.5772/57552

Cited by 23 publications

(38 citation statements)

References 124 publications

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“…Interestingly, the three hemocyanins induced significant elevations in the mRNA levels of CD40L (Cd40lg) in macrophages, which might support the observed nonspecific antitumor activity of hemocyanins in several cancers (6,71). CD40-CD40L interactions promote the activation of NK cells by murine macrophages (72).…”

Section: Discussionmentioning

confidence: 84%

“…The aim of this study was to determine how mollusk hemocyanins, in the absence of additional adjuvants, trigger an innate immune response that leads to the maturation of a Th1-specific adaptive immune response together with powerful nonspecific immunomodulatory effects, including an antitumor capacity in certain cancers (3,5,6). The immune response is triggered by a proinflammatory signal that arises from various components of the innate immune system that sense invading pathogens (52) or danger/alarm signals from injured cells (53), phenomena in which phagocytic cells and APCs are fundamental.…”

Section: Discussionmentioning

confidence: 99%

“…Several authors have explained that the immunomodulatory effect of hemocyanins is a result of the extremely complex xenogeneic structures (4,6,21), which resemble an icosahedral virus capsid (22). In fact, mollusk hemocyanins are enormous glycoproteins of ∼4 MDa, composed by an intricate arrangement of non-covalently linked subunits.…”

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confidence: 99%

“…However, the biodiversity of hemocyanins has prompted interest in developing new candidates with better biochemical and immunological properties because the supply depends on natural resources. Therefore, hemocyanins from Concholepas concholepas (CCH) (7) and Fissurella latimarginata (FLH) (8), among others (6), have been studied. CCH is used as a carrier (9) and experimental Ag (10), and it has been preclinically evaluated in a murine model of superficial bladder cancer and showed antitumor effects similar to KLH (11).…”

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confidence: 99%

“…Therefore, hemocyanins have been used as a biotechnological tool in the development of Abs and vaccines, and also as immunomodulators such as nonspecific immunostimulants in superficial bladder cancer (3). The hemocyanin purified from the giant keyhole limpet gastropod Megathura crenulata, commonly known as keyhole limpet hemocyanin (KLH), has traditionally been used for those purposes (4)(5)(6). However, the biodiversity of hemocyanins has prompted interest in developing new candidates with better biochemical and immunological properties because the supply depends on natural resources.…”

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confidence: 99%

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Hemocyanins Stimulate Innate Immunity by Inducing Different Temporal Patterns of Proinflammatory Cytokine Expression in Macrophages

Zhong

Arancibia

Born

et al. 2016

The Journal of Immunology

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Hemocyanins induce a potent Th1-dominant immune response with beneficial clinical outcomes when used as a carrier/adjuvant in vaccines and nonspecific immunostimulant in cancer. However, the mechanisms by which hemocyanins trigger innate immune responses, leading to beneficial adaptive immune responses, are unknown. This response is triggered by a proinflammatory signal from various components, of which macrophages are an essential part. To understand how these proteins influence macrophage response, we investigated the effects of mollusks hemocyanins with varying structural and immunological properties, including hemocyanins from Concholepas concholepas, Fissurella latimarginata, and Megathura crenulata (keyhole limpet hemocyanin), on cultures of peritoneal macrophages. Hemocyanins were phagocytosed and slowly processed. Analysis of this process showed differential gene expression along with protein levels of proinflammatory markers, including IL-1β, IL-6, IL-12p40, and TNF-α. An extended expression analysis of 84 cytokines during a 24-h period showed a robust proinflammatory response for F. latimarginata hemocyanin in comparison with keyhole limpet hemocyanin and C. concholepas hemocyanin, which was characterized by an increase in the transcript levels of M1 cytokines involved in leukocyte recruitment. These cytokine genes included chemokines (Cxcl1, Cxcl3, Cxcl5, Ccl2, and Ccl3), ILs (Il1b and Ifng), growth factors (Csf2 and Csf3), and TNF family members (Cd40lg). The protein levels of certain cytokines were increased. However, every hemocyanin maintains downregulated key M2 cytokine genes, including Il4 and Il5. Collectively, our data demonstrate that hemocyanins are able to trigger the release of proinflammatory factors with different patterns of cytokine expression, suggesting differential signaling pathways and transcriptional network mechanisms that lead to the activation of M1-polarized macrophages.

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Section: Discussionmentioning

confidence: 84%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Hemocyanins Stimulate Innate Immunity by Inducing Different Temporal Patterns of Proinflammatory Cytokine Expression in Macrophages

Zhong

Arancibia

Born

et al. 2016

The Journal of Immunology

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C‐type lectin receptors MR and DC‐SIGN are involved in recognition of hemocyanins, shaping their immunostimulatory effects on human dendritic cells

et al. 2021

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Hemocyanins are used as immunomodulators in clinical applications because they induce a strong Th1‐biased cell‐mediated immunity, which has beneficial effects. They are multiligand glycosylated molecules with abundant and complex mannose‐rich structures. It remains unclear whether these structures influence hemocyanin‐induced immunostimulatory processes in human APCs. We have previously shown that hemocyanin glycans from Concholepas concholepas (CCH), Fissurella latimarginata (FLH), and Megathura crenulata (KLH), participate in their immune recognition and immunogenicity in mice, interacting with murine C‐type lectin receptors (CLRs). Here, we studied the interactions of these hemocyanins with two major mannose‐binding CLRs on monocyte‐derived human DCs: MR (mannose receptor) and DC‐SIGN (DC‐specific ICAM‐3–grabbing nonintegrin). Diverse analyses showed that hemocyanins are internalized by a mannose‐sensitive mechanism. This process was calcium dependent. Moreover, hemocyanins colocalized with MR and DC‐SIGN, and were partly internalized through clathrin‐mediated endocytosis. The hemocyanin‐mediated proinflammatory cytokine response was impaired when using deglycosylated FLH and KLH compared to CCH. We further showed that hemocyanins bind to human MR and DC‐SIGN in a carbohydrate‐dependent manner with affinity constants in the physiological concentration range. Overall, we showed that these three clinically valuable hemocyanins interact with human mannose‐sensitive CLRs, initiating an immune response and promoting a Th1 cell‐driving potential.

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Molluscan hemocyanin: structure, evolution, and physiology

et al. 2017

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Most molluscs have blue blood because their respiratory molecule is hemocyanin, a type-3 copper-binding protein that turns blue upon oxygen binding. Molluscan hemocyanins are huge cylindrical multimeric glycoproteins that are found freely dissolved in the hemolymph. With molecular masses ranging from 3.3 to 13.5 MDa, molluscan hemocyanins are among the largest known proteins. They form decamers or multi-decamers of 330- to 550-kDa subunits comprising more than seven paralogous functional units. Based on the organization of functional domains, they assemble to form decamers, di-decamers, and tri-decamers. Their structure has been investigated using a combination of single particle electron cryo-microsopy of the entire structure and high-resolution X-ray crystallography of the functional unit, although, the one exception is squid hemocyanin for which a crystal structure analysis of the entire molecule has been carried out. In this review, we explain the molecular characteristics of molluscan hemocyanin mainly from the structural viewpoint, in which the structure of the functional unit, architecture of the huge cylindrical multimer, relationship between the composition of the functional unit and entire tertiary structure, and possible functions of the carbohydrates are introduced. We also discuss the evolutionary implications and physiological significance of molluscan hemocyanin.

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Mollusk Hemocyanins as Natural Immunostimulants in Biomedical Applications

Cited by 23 publications

References 124 publications

Hemocyanins Stimulate Innate Immunity by Inducing Different Temporal Patterns of Proinflammatory Cytokine Expression in Macrophages

Hemocyanins Stimulate Innate Immunity by Inducing Different Temporal Patterns of Proinflammatory Cytokine Expression in Macrophages

C‐type lectin receptors MR and DC‐SIGN are involved in recognition of hemocyanins, shaping their immunostimulatory effects on human dendritic cells

Molluscan hemocyanin: structure, evolution, and physiology

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