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Developmental and Comparative Immunology May 2024Microbial drug resistance is becoming increasingly severe due to antibiotic abuse. The development and utilization of antimicrobial peptides is one of the important ways...
Microbial drug resistance is becoming increasingly severe due to antibiotic abuse. The development and utilization of antimicrobial peptides is one of the important ways to solve this difficult problem. Crustins are a family of antimicrobial peptides that play important roles in the innate immune system of crustaceans. Several types of crustins exist in shrimp and their activities vary greatly. In the present study, we studied the immune function of one newly identified crustin and found that the type VI crustin encoding gene in Litopenaeus vannamei (LvCrustinVI) was mainly expressed in gills. Its expression was significantly up-regulated after Vibrio parahaemolyticus infection and knockdown of the gene promoted Vibrio proliferation in the hepatopancreas of shrimp, indicating that LvCrustinVI was involved in pathogens infection. The recombinant LvCrustinVI (rLvCrustinVI) showed strong inhibitory activities against both Gram-negative and Gram-positive bacteria, and exhibited binding activities with the bacteria and bacterial polysaccharides including Glu, LPS and PGN. In the presence of Ca, rLvCrustinVI showed a strong agglutination effect on V. parahaemolyticus and could significantly enhance the phagocytic ability of shrimp hemocytes against V. parahaemolyticus. In conclusion, LvCrustinVI played important roles as antimicrobial peptide and opsonin in the innate immune defense of L. vannamei. The study enriched our understanding of the functional activity of Crustin and provides an important basis for the development and utilization of antimicrobial peptides.
Topics: Animals; Vibrio parahaemolyticus; Antimicrobial Peptides; Immunity, Innate; Anti-Bacterial Agents; Vibrio Infections; Penaeidae; Arthropod Proteins; Phylogeny
PubMed: 38316231
DOI: 10.1016/j.dci.2024.105146 -
Fish & Shellfish Immunology Feb 2024C-type lectins (CTLs) function as pattern recognition receptors (PRRs) by recognizing invading microorganisms, thereby triggering downstream immune events against...
C-type lectins (CTLs) function as pattern recognition receptors (PRRs) by recognizing invading microorganisms, thereby triggering downstream immune events against infected pathogens. In this study, a novel CTL containing a low-density lipoprotein receptor class A (LDLa) domain was obtained from Litopenaeus vannamei, designed as LvLDLalec. Stimulation by the bacterial pathogen Vibrio anguillarum (V. anguillarum) resulted in remarkable up-regulation of LvLDLalec, as well as release of LvLDLalec into hemolymph. The rLvLDLalec protein possessed broad-spectrum bacterial binding and agglutinating activities, as well as hemocyte attachment ability. Importantly, LvLDLalec facilitated the bacterial clearance in shrimp hemolymph and protected shrimp from bacterial infection. Further studies revealed that LvLDLalec promoted hemocytes phagocytosis against V. anguillarum and lysosomes were involved in the process. Meanwhile, LvLDLalec participated in humoral immunity through activating and inducing nuclear translocation of Dorsal to regulate phagocytosis-related genes and antimicrobial peptides (AMPs) genes, thereby accelerated the removal of invading pathogens in vivo and improved the survival rate of L. vannamei. These results unveil that LvLDLalec serves as a PRR participate in cellular and humoral immunity exerting opsonin activity to play vital roles in the immune regulatory system of L. vannamei.
Topics: Animals; Lectins, C-Type; Phagocytosis; Bacterial Infections; Receptors, Pattern Recognition; Bacteria; Crustacea; Penaeidae; Immunity, Innate; Hemocytes; Arthropod Proteins
PubMed: 38185393
DOI: 10.1016/j.fsi.2024.109361 -
Cells & Development Mar 2024Epithelial outpocketing, tunic softening, mesenchymal cell death, dedifferentiation/transdifferentiation, and resistance to environmental stress are major events that...
Epithelial outpocketing, tunic softening, mesenchymal cell death, dedifferentiation/transdifferentiation, and resistance to environmental stress are major events that occur during asexual reproduction by budding in the tunicate, Polyandrocarpa misakiensis. To identify the molecules underlying these events and compare them with those operating in regeneration, differential gene expression profiles were developed in buds and zooids. Among approximately 40,000 contigs, 21 genes were identified as potentially being involved in asexual reproduction. Genes related to tunic softening, phagocytosis-stimulating opsonin, and stress resistance were activated in the very early stage of budding. At the later stage of budding when buds separated from the parent and entered the developmental stage, genes for cell adhesion, cell death, and differentiation were activated. The transcription factor AP2 was spatio-temporally expressed in a similar pattern to the tunic-softening gene endoglucanase (EndoG). AP2 mRNA activated EndoG when introduced into zooids by electroporation. Eight out of 21 budding-related genes were significantly activated by AP2 mRNA. Polyandrocarpa zooids possess regenerative potential other than budding. Zooidal regeneration accompanied cell death/phagocytosis, cell-cell adhesion/communication, and dedifferentiation/redifferentiation. Consistent with morphological features, eight related genes including SP8 transcription factor were activated during zooidal regeneration. Most of these genes were identical to those induced by AP2 mRNA, indicating that asexual reproduction in P. misakiensis shares AP2-regulated downstream genes with zooidal regeneration. The present results suggest that SP8 may be indispensable for both budding and regeneration and that the potential dedifferentiation-related gene SOXB1 plays a minor role in zooidal regeneration.
Topics: Animals; Transcription Factor AP-2; Urochordata; Reproduction, Asexual; Cell Differentiation; RNA, Messenger
PubMed: 38007002
DOI: 10.1016/j.cdev.2023.203885 -
Cell Biochemistry and Function Dec 2023Opsonization plays a pivotal role in hindering controlled drug release from nanoformulations due to macrophage-mediated nanoparticle destruction. While first and... (Review)
Review
Opsonization plays a pivotal role in hindering controlled drug release from nanoformulations due to macrophage-mediated nanoparticle destruction. While first and second-generation delivery systems, such as lipoplexes (50-150 nm) and quantum dots, hold immense potential in revolutionizing disease treatment through spatiotemporal controlled drug delivery, their therapeutic efficacy is restricted by the selective labeling of nanoparticles for uptake by reticuloendothelial system and mononuclear phagocyte system via various molecular forces, such as electrostatic, hydrophobic, and van der Waals bonds. This review article presents novel insights into surface-modification techniques utilizing macromolecule-mediated approaches, including PEGylation, di-block copolymerization, and multi-block polymerization. These techniques induce stealth properties by generating steric forces to repel micromolecular-opsonins, such as fibrinogen, thereby mitigating opsonization effects. Moreover, advanced biological methods, like cellular hitchhiking and dysopsonic protein adsorption, are highlighted for their potential to induce biological camouflage by adsorbing onto the nanoparticulate surface, leading to immune escape. These significant findings pave the way for the development of long-circulating next-generation nanoplatforms capable of delivering superior therapy to patients. Future integration of artificial intelligence-based algorithms, integrated with nanoparticle properties such as shape, size, and surface chemistry, can aid in elucidating nanoparticulate-surface morphology and predicting interactions with the immune system, providing valuable insights into the probable path of opsonization.
Topics: Humans; Polyethylene Glycols; Opsonization; Artificial Intelligence; Drug Delivery Systems; Opsonin Proteins; Nanoparticles
PubMed: 37933222
DOI: 10.1002/cbf.3880 -
Frontiers in Cellular and Infection... 2023The complement receptor CR3, also known as integrin Mac-1 (CD11b/CD18), is one of the major phagocytic receptors on the surface of neutrophils and macrophages. We...
The complement receptor CR3, also known as integrin Mac-1 (CD11b/CD18), is one of the major phagocytic receptors on the surface of neutrophils and macrophages. We previously demonstrated that in its protein ligands, Mac-1 binds sequences enriched in basic and hydrophobic residues and strongly disfavors negatively charged sequences. The avoidance by Mac-1 of negatively charged surfaces suggests that the bacterial wall and bacterial capsule possessing net negative electrostatic charge may repel Mac-1 and that the cationic Mac-1 ligands can overcome this evasion by acting as opsonins. Indeed, we previously showed that opsonization of Gram-negative with several cationic peptides, including PF4 (Platelet Factor 4), strongly augmented phagocytosis by macrophages. Here, we investigated the effect of recombinant PF4 (rPF4) on phagocytosis of Gram-positive and examined its impact in a mouse model of peritonitis. Characterization of the interaction of rPF4 with nonencapsulated and encapsulated showed that rPF4 localizes on the bacterial surface, thus making it available for Mac-1. Furthermore, rPF4 did not have direct bactericidal and bacteriostatic activity and was not toxic to host cells. rPF4 enhanced phagocytosis of bioparticles by various primary and cultured Mac-1-expressing leukocytes by several folds. It also increased phagocytosis of live nonencapsulated and encapsulated bacteria. Notably, the augmentation of phagocytosis by rPF4 did not compromise the intracellular killing of by macrophages. Using a murine peritonitis model, we showed that treatment of infected mice with rPF4 caused a significant increase in the clearance of antibiotic-susceptible and its methicillin-resistant (MRSA) variant and markedly improved survival. These findings indicate that rPF4 binding to the bacterial surface circumvents its antiphagocytic properties, improving host defense against antibiotic-susceptible and antibiotic-resistant bacteria.
Topics: Animals; Mice; Methicillin-Resistant Staphylococcus aureus; Anti-Bacterial Agents; Platelet Factor 4; Staphylococcus aureus; Disease Models, Animal; Phagocytosis; Macrophage-1 Antigen; Immunologic Factors; Peritonitis
PubMed: 37868353
DOI: 10.3389/fcimb.2023.1217103 -
Nanoscale Nov 2023The biological behavior and fate of nanoparticles are dependent on their retention time in the blood circulation system. The protein corona components, especially...
The biological behavior and fate of nanoparticles are dependent on their retention time in the blood circulation system. The protein corona components, especially opsonins, and dysopsonins, adsorbed on the nanoparticle surface determine their blood circulation time. The protein corona formation is a dynamic process that involves the competition between different proteins to be adsorbed on the nanoparticles. Therefore, studying how proteins compete and are oriented on the nanoparticle surface is essential. We hypothesized that the presence of opsonins (immunoglobulin (IgG)) might affect the adsorption of dysopsonins (human serum albumin (HSA)) and . Using the molecular dynamics simulations, we showed that the adsorption of HSA on the GO surface after the IgG adsorption is more probable than the opposite order of adsorption. It was also observed that the higher lateral diffusion of the HSA compared to the IgG helped the system reach a more stable configuration while the initial adsorption of the HSA limits the lateral diffusion of IgG. Therefore, replacing IgG adsorbed on the GO surface with HSA is plausible while the reverse process is less likely to occur. This study revealed that albumin might extend the blood circulation time of GO by replacing opsonins (IgG).
Topics: Humans; Opsonin Proteins; Protein Corona; Nanoparticles; Serum Albumin, Human; Immunoglobulin G; Adsorption
PubMed: 37860936
DOI: 10.1039/d3nr03823h -
Frontiers in Immunology 2023C-reactive protein (CRP) is one of the major members of the family of acute phase proteins (APP). Interest in this CRP was the result of a seminal discovery of its... (Review)
Review
C-reactive protein (CRP) is one of the major members of the family of acute phase proteins (APP). Interest in this CRP was the result of a seminal discovery of its pattern of response to pneumococcal infection in humans. CRP has the unique property of reacting with phosphocholine-containing substances, such as pneumococcal C-polysaccharide, in the presence of Ca. The attention regarding the origin of CRP and its multifunctionality has gripped researchers for several decades. The reason can be traced to the integrated evolution of CRP in the animal kingdom. CRP has been unequivocally listed as a key indicator of infectious and inflammatory diseases including autoimmune diseases. The first occurrence of CRP in the evolutionary ladder appeared in arthropods followed by molluscs and much later in the chordates. The biological significance of CRP has been established in the animal kingdom starting from invertebrates. Interestingly, the site of synthesis of CRP is mainly the liver in vertebrates, while in invertebrates it is located in diverse tissues. CRP is a multifunctional player in the scenario of innate immunity. CRP acts as an opsonin in the area of complement activation and phagocytosis. Interestingly, CRP upregulates and downregulates both cytokine production and chemotaxis. Considering various studies of CRP in humans and non-human animals, it has been logically proposed that CRP plays a common role in animals. CRP also interacts with Fcγ receptors and triggers the inflammatory response of macrophages. CRP in other animals such as primates, fish, echinoderms, arthropods, and molluscs has also been studied in some detail which establishes the evolutionary significance of CRP. In mammals, the increase in CRP levels is an induced response to inflammation or trauma; interestingly, in arthropods and molluscs, CRP is constitutively expressed and represents a major component of their hemolymph. Investigations into the primary structure of CRP from various species revealed the overall relatedness between vertebrate and invertebrate CRP. Invertebrates lack an acquired immune response; they are therefore dependent on the multifunctional role of CRP leading to the evolutionary success of the invertebrate phyla.
Topics: Animals; C-Reactive Protein; Inflammation; Invertebrates; Mammals; Opsonin Proteins; Phagocytosis; Humans
PubMed: 37860004
DOI: 10.3389/fimmu.2023.1238411 -
Frontiers in Immunology 2023Systemic amyloidosis is a progressive disorder characterized by the extracellular deposition of amyloid fibrils and accessory proteins in visceral organs and tissues....
INTRODUCTION
Systemic amyloidosis is a progressive disorder characterized by the extracellular deposition of amyloid fibrils and accessory proteins in visceral organs and tissues. Amyloid accumulation causes organ dysfunction and is not generally cleared by the immune system. Current treatment focuses on reducing amyloid precursor protein synthesis and slowing amyloid deposition. However, curative interventions will likely also require removal of preexisting amyloid deposits to restore organ function. Here we describe a prototypic pan-amyloid binding peptide-antibody fusion molecule (mIgp5) that enhances macrophage uptake of amyloid.
METHODS
The murine IgG1-IgG2a hybrid immunoglobulin with a pan amyloid-reactive peptide, p5, fused genetically to the N-terminal of the immunoglobulin light chain was synthesized in HEK293T/17 cells. The binding of the p5 peptide moiety was assayed using synthetic amyloid-like fibrils, human amyloid extracts and amyloid-laden tissues as substrates. Binding of radioiodinated mIgp5 with amyloid deposits was evaluated in a murine model of AA amyloidosis using small animal imaging and microautoradiography. The bioactivity of mIgp5 was assessed in complement fixation and phagocytosis assays in the presence of patient-derived amyloid extracts and synthetic amyloid fibrils as substrates and in the presence or absence of human serum.
RESULTS
Murine Igp5 exhibited highly potent binding to AL and ATTR amyloid extracts and diverse types of amyloid in formalin-fixed tissue sections. In the murine model of systemic AA amyloidosis, I-mIgp5 bound rapidly and specifically to amyloid deposits in all organs, including the heart, with no evidence of non-specific uptake in healthy tissues. The bioactivity of the immunoglobulin Fc domain was uncompromised in the context of mIgp5 and served as an effective opsonin. Macrophage-mediated uptake of amyloid extract and purified amyloid fibrils was enhanced by the addition of mIgp5. This effect was exaggerated in the presence of human serum coincident with deposition of complement C5b9.
CONCLUSION
Immunostimulatory, amyloid-clearing therapeutics can be developed by incorporating pan-amyloid-reactive peptides, such as p5, as a targeting moiety. The immunologic functionality of the IgG remains intact in the context of the fusion protein. These data highlight the potential use of peptide-antibody fusions as therapeutics for all types of systemic amyloidosis.
Topics: Mice; Animals; Humans; Disease Models, Animal; HEK293 Cells; Plaque, Amyloid; Amyloidosis; Amyloid; Amyloidogenic Proteins; Peptides; Immunoglobulin Light Chains
PubMed: 37854603
DOI: 10.3389/fimmu.2023.1275372 -
Acta Pharmacologica Sinica Mar 2024Higher drug loading employed in nanoscale delivery platforms is a goal that researchers have long sought after. But such viewpoint remains controversial because the...
Higher drug loading employed in nanoscale delivery platforms is a goal that researchers have long sought after. But such viewpoint remains controversial because the impacts that nanocarriers bring about on bodies have been seriously overlooked. In the present study we investigated the effects of drug loading on the in vivo performance of PEGylated liposomal doxorubicin (PLD). We prepared PLDs with two different drug loading rates: high drug loading rate, H-Dox, 12.9% w/w Dox/HSPC; low drug loading rate, L-Dox, 2.4% w/w Dox/HSPC (L-Dox had about 5 folds drug carriers of H-Dox at the same Dox dose). The pharmaceutical properties and biological effects of H-Dox and L-Dox were compared in mice, rats or 4T1 subcutaneous tumor-bearing mice. We showed that the lowering of doxorubicin loading did not cause substantial shifts to the pharmaceutical properties of PLDs such as in vitro and in vivo stability (stable), anti-tumor effect (equivalent effective), as well as tissue and cellular distribution. Moreover, it was even more beneficial for mitigating the undesired biological effects caused by PLDs, through prolonging blood circulation and alleviating cutaneous accumulation in the presence of pre-existing anti-PEG Abs due to less opsonins (e.g. IgM and C3) deposition on per particle. Our results warn that the effects of drug loading would be much more convoluted than expected due to the complex intermediation between nanocarriers and bodies, urging independent investigation for each individual delivery platform to facilitate clinical translation and application.
Topics: Mice; Rats; Animals; Cell Line, Tumor; Doxorubicin; Polyethylene Glycols; Drug Carriers
PubMed: 37845342
DOI: 10.1038/s41401-023-01169-5 -
Microbiological Research Dec 2023The human complement system is an important part of the innate immune response in the fight against invasive bacteria. Complement responses can be activated... (Review)
Review
The human complement system is an important part of the innate immune response in the fight against invasive bacteria. Complement responses can be activated independently by the classical pathway, the lectin pathway, or the alternative pathway, each resulting in the formation of a C3 convertase that produces the anaphylatoxin C3a and the opsonin C3b by specifically cutting C3. Other important features of complement are the production of the chemotactic C5a peptide and the generation of the membrane attack complex to lyse intruding pathogens. Invasive pathogens like Staphylococcus aureus and several species of the genus Streptococcus have developed a variety of complement evasion strategies to resist complement activity thereby increasing their virulence and potential to cause disease. In this review, we focus on secreted complement evasion factors that assist the bacteria to avoid opsonization and terminal pathway lysis. We also briefly discuss the potential role of complement evasion factors for the development of vaccines and therapeutic interventions.
Topics: Humans; Gram-Positive Cocci; Immunity, Innate; Staphylococcal Infections; Immune System; Immune Evasion
PubMed: 37826985
DOI: 10.1016/j.micres.2023.127512