Molecular Identification and Agglutination Characteristics of C-Type Lectin Receptor in Pacific White Shrimp Litopenaeus vannamei
ZHANG Huan, WANG Yaohua, YAN Maocang, HUANG Xianke, LUO Kui, LI Fang, ZHANG Min
Wenzhou Key Laboratory of Marine Biological Genetics and Breeding, Zhejiang Key Laboratory of Exploitation and Preservation of Coastal Bioresource, Zhejiang Mariculture Research Institute, Wenzhou 325000, China
Abstract:In order to probe into the biological function of C-type lectin, the C-type lectin LvCTLP1 gene in hepatopancreas of Pacific white shrimp Litopenaeus vannamei with body weight of 20.328—22.478 g and body length of 18.2—20.6 cm was obtained by PCR, and the full-length sequence of the genetic gene was analyzed and its sequence characteristics were analyzed, and the tissue distribution of LvCTLP1 gene and the bacterial agglutination characteristics of the recombinant protein were analyzed by fluorescence quantitative real-time PCR. Bioinformatics analysis showed that the LvCTLP1 gene encoded a 723 bp cDNA sequence, containing a 459 bp ORF that translates into a 152-amino acid polypeptide. LvCTLP1 features an N-terminal signal peptide (21 amino acids) and a C-terminal carbohydrate recognition domain (124 amino acids). The LvCTLP1 gene was shown to be the maximal expressed level in the hepatopancreas, with Ca2+-dependent bacterial agglutination activity in the recombinant rLvCTLP1 exhibited, specifically against Escherichia coli and Vibrio anguillarum. The agglutination activity of rLvCTLP1 against E. coli was found to be significantly inhibited by multiple saccharides, including D-glucose, D-galactose, D-mannose, D-fructose, maltose, sucrose, and alginose, indicating specific carbohydrate -binding capability of this lectin. These findings suggest that LvCTLP1 may function as a pattern recognition receptor capable of identifying bacterial pathogens through carbohydrate-PAMP interactions, thereby participating in antibacterial immune response ofPacific white shrimp.
[1] 张伟权.世界重要养殖品种——南美白对虾生物学简介[J].海洋科学,1990,14(3):69-73. [2] 王彩理,毕国栋,于爱美,等.凡纳滨对虾的营养功效和安全利用[J].农产品加工,2021(24):72-74. [3] 农业农村部渔业渔政管理局,全国水产技术推广总站,中国水产学会.2023中国渔业统计年鉴[M].北京:中国农业出版社,2023. [4] IRANI M, RAJABI ISLAMI H, NAFISI BAHABADI M, et al. Production of Pacific white shrimp under different stocking density in a zero-water exchange biofloc system:effects on water quality, zootechnical performance, and body composition[J]. Aquacultural Engineering,2023,100:102313. [5] CHEN J D, YANG F, CHENG T, et al. Glutathione-rich yeast hydrolysate makes the contributions to growth performance, healthy of Pacific white shrimp (Litopenaeus vannamei), and helps shrimp resist nitrite stress[J]. Aquaculture Reports,2023,33:101825. [6] LOW C F, CHONG C M. Peculiarities of innate immune memory in crustaceans[J]. Fish & Shellfish Immunology,2020,104:605-612. [7] BUCHMANN K. Evolution of innate immunity:clues from invertebrates via fish to mammals[J]. Frontiers in Immunology,2014,5:459. [8] MEDZHITOV R, JANEWAY C A Jr. Decoding the patterns of self and nonself by the innate immune system[J]. Science,2002,296(5566):298-300. [9] SÁNCHEZ-SALGADO J L, PEREYRA M A, ALPUCHE-OSORNO J J, et al. Pattern recognition receptors in the crustacean immune response against bacterial infections[J]. Aquaculture,2021,532:735998. [10] ROY S, BOSSIER P, NOROUZITALLAB P, et al. Trained immunity and perspectives for shrimp aquaculture[J]. Reviews in Aquaculture,2020,12(4):2351-2370. [11] GEIJTENBEEK T B H, GRINGHUIS S I. Signalling through C-type lectin receptors:shaping immune responses[J]. Nature Reviews. Immunology,2009,9(7):465-479. [12] VASTA G R, NITA-LAZAR M, GIOMARELLI B, et al. Structural and functional diversity of the lectin repertoire in teleost fish: relevance to innate and adaptive immunity[J]. Developmental & Comparative Immunology,2011,35(12):1388-1399. [13] DRICKAMER K, TAYLOR M E. Recent insights into structures and functions of C-type lectins in the immune system[J]. Current Opinion in Structural Biology,2015,34:26-34. [14] 李红岩,张祥敏,杨帅奇.海洋生物C型凝集素研究进展[J].中国海洋大学学报(自然科学版),2020,50(9):113-119. [15] CAMBI A, KOOPMAN M, FIGDOR C G. How C-type lectins detect pathogens[J]. Cellular Microbiology,2005,7(4):481-488. [16] ZELENSKY A N, GREADY J E. The C-type lectin-like domain superfamily[J]. The FEBS Journal,2005,272(24):6179-6217. [17] ZELENSKY A, GREADY J. Analysis of whole-genome and other data resources to characterize the molecular, structural, and evolutionary diversity of C-type lectins and discover new genes[M]//VASTA G R, AHMED H. Animal Lectins. Boca Raton:CRC Press,2008. [18] BI J X, NING M X, XIE X J, et al. A typical C-type lectin, perlucin-like protein, is involved in the innate immune defense of whiteleg shrimp Litopenaeus vannamei[J]. Fish & Shellfish Immunology,2020,103:293-301. [19] DENIS M, THAYPPAN K, RAMASAMY S, et al. Lectin in innate immunity of Crustacea[J]. Austin Biology,2016,1(1):1001. [20] WANG X W, WANG J X. Diversity and multiple functions of lectins in shrimp immunity[J]. Developmental & Comparative Immunology,2013,39(1/2):27-38. [21] LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2 (-Delta Delta C(T)) Method[J]. Methods,2001,25(4):402-408. [22] DAY A J. The C-type carbohydrate recognition domain (CRD) superfamily[J]. Biochemical Society Transactions,1994,22(1):83-88. [23] DAMBUZA I M, BROWN G D. C-type lectins in immunity:recent developments[J]. Current Opinion in Immunology,2015,32:21-27. [24] 罗展,张继泉,李富花,等.凡纳滨对虾C-型凝集素LvLec2对不同刺激的免疫应答[J].海洋科学,2010,34(11):103-110. [25] 秦玉凯.两种斑节对虾C型凝集素抗菌功能研究及耐低盐、耐氨氮等相关表达模式分析[D].上海:上海海洋大学,2019. [26] BALIARSINGH S, SAHOO S, JO Y H, et al. Molecular cloning, sequence characterization, and expression analysis of C-type lectin (CTL) and ER-Golgi intermediate compartment 53-kDa protein (ERGIC-53) homologs from the freshwater prawn, Macrobrachium rosenbergii[J]. Aquaculture International,2022,30(2):1011-1035. [27] LUO T, ZHANG X B, SHAO Z Z, et al. PmAV, a novel gene involved in virus resistance of shrimp Penaeus monodon1[J]. FEBS Letters,2003,551(1/2/3):53-57. [28] ZHANG Y, QIU L M, SONG L S, et al. Cloning and characterization of a novel C-type lectin gene from shrimp Litopenaeus vannamei[J]. Fish & Shellfish Immunology,2009,26(1):183-192. [29] COSTA F F, VALENÇA N S, SILVA A P, et al. Cloning and molecular modeling of Litopenaeus vannamei (Penaeidae) C-type lectin homologs with mutated mannose binding domain-2[J]. Genetics and Molecular Research,2011,10(2):650-664. [30] WEI X M, LIU X Q, YANG J M, et al. Two C-type lectins from shrimp Litopenaeus vannamei that might be involved in immune response against bacteria and virus[J]. Fish & Shellfish Immunology,2012,32(1):132-140. [31] MA T H T, TIU S H K, HE J G, et al. Molecular cloning of a C-type lectin (LvLT) from the shrimp Litopenaeus vannamei:early gene down-regulation after WSSV infection[J]. Fish & Shellfish Immunology,2007,23(2):430-437. [32] ZHAO Z Y, YIN Z X, XU X P, et al. A novel C-type lectin from the shrimp Litopenaeus vannamei possesses anti-white spot syndrome virus activity[J]. Journal of Virology,2009,83(1):347-356. [33] SONG F, CHEN G L, LU K C, et al. Identification and functional characterization of a C-type lectin gene from Litopenaeus vannamei that is associated with ER-stress response[J]. Fish & Shellfish Immunology,2019,93:977-985. [34] YU X Q, KANOST M R. Immulectin-2, a lipopolysaccharide-specific lectin from an insect, Manduca sexta, is induced in response to gram-negative bacteria[J]. Journal of Biological Chemistry,2000,275(48):37373-37381. [35] 郝少彦,尹丝雨,龚晨,等.一种凡纳滨对虾新的C型凝集素基因(LvLc2)的克隆及免疫应答特征[J].水产学报,2021,45(6):948-957. [36] HUANG Y, AN L, HUI K M, et al. An LDLa domain-containing C-type lectin is involved in the innate immunity of Eriocheir sinensis[J]. Developmental & Comparative Immunology,2014,42(2):333-344. [37] LUO T, YANG H J, LI F, et al. Purification, characterization and cDNA cloning of a novel lipopolysaccharide-binding lectin from the shrimp Penaeus monodon[J]. Developmental & Comparative Immunology,2006,30(7):607-617. [38] HUANG X, HUANG Y, SHI Y R, et al. Function of a novel C-type lectin with two CRD domains from Macrobrachium rosenbergii in innate immunity[J]. Developmental & Comparative Immunology,2015,49(1):121-126. [39] LI Y B, PAN L Q, YU J H. The injection of one recombinant C-type lectin (LvLec) induced the immune response of hemocytes in Litopenaeus vannamei[J]. Fish & Shellfish Immunology,2022,124:324-331.