Next Article in Journal
Epidemiology of Tick-Borne Encephalitis (TBE) in Germany, 2001–2018
Previous Article in Journal
Candida albicans at Host Barrier Sites: Pattern Recognition Receptors and Beyond
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Pathogens
Volume 8
Issue 1
10.3390/pathogens8010041
pathogens-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles thumb_up
...
Endorse textsms
...
Comment
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

The Genetic Polymorphisms of 24 Base Pair Duplication and Point G102S of Human Chitotriosidase to Bancroftian Filariasis at the Thai–Myanmar Border

by
Vivornpun Sanprasert
1,2,
Sarit Charuchaibovorn
1 and
Surang Nuchprayoon
1,2,*
1
Lymphatic Filariasis and Tropical Medicine Research Unit, Chulalongkorn Medical Research Center, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand
2
Department of Parasitology, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand
*
Author to whom correspondence should be addressed.
Pathogens 2019, 8(1), 41; https://doi.org/10.3390/pathogens8010041
Submission received: 7 February 2019 / Revised: 15 March 2019 / Accepted: 20 March 2019 / Published: 25 March 2019
(This article belongs to the Section Immunological Responses and Immune Defense Mechanisms)
Browse Figure
Versions Notes

Abstract

:
Lymphatic filariasis, caused by lymphatic filarial parasites, Wuchereria bancrofti, and Brugia malayi, causes significant morbidity and disability to 120 million people in the tropics and subtropics. Chitin has an important role for embryogenesis in adult worms and is a component of microfilaria sheath. Human chitotriosidase (CHIT1) is a chitin-degrading enzyme which provides a protective role against chitin-containing pathogens. Here, we determined the association of CHIT1 polymorphisms with susceptibility to bancroftian filariasis (BF) in 88 individuals at the Thai–Myanmar border. Two common polymorphisms of CHIT1, contributing inactive CHIT protein, including 24 base pair (24 bp) duplication in exon 10, and p. G102S in exon 4 were genotyped by allele-specific Polymerase Chain Reaction (PCR) and PCR sequencing, respectively. Unexpectedly, genotype frequencies of 24 bp duplication insertion homozygous (INS/INS) were significantly higher in endemic normal (EN) (40.0%) than BF patients (31.4%). In contrast, genotype frequencies of p. G102S homozygous (A/A) in BF patients (21.6%) was higher than in EN (19.0%) without statistical difference. Mutant allele frequencies of 24 bp duplication were 0.6125 (98/160) and p. G102S were 0.392 (69/176). Genotype and allele frequencies of CHIT1, 24 bp duplication, and p. G102S, showed no association with BF patients.

1. Introduction

Lymphatic filariasis is considered as a neglected tropical disease (NTD) targeted to eliminate by 2020 [1]. The disease is a mosquito-borne disease caused by lymphatic filarial parasites, including Wuchereria bancrofti, Brugia malayi, and B. timori. Over 120 million people in tropic and subtropic countries are infected with these parasites [2]. Chronic infection with mosquito-transmitted filarial worms leads to lymphatic dysfunction, resulting in enlargement of body parts, causing pain, severe disability, and social stigma. There are two species of lymphatic filarial parasites in Thailand, i.e., W. bancrofti (at the Thai–Myanmar border), and B. malayi (at the Thai–Malaysia border) [2].
The differential susceptibility of lymphatic filariasis is affected by socioeconomic status, personal activities, age, gender, and host genetics [2,3,4]. Several studies have demonstrated the association of polymorphisms of host defense pathway genes with susceptibility and clinical outcomes of bancroftian filariasis (BF). Polymorphisms of mannose-binding lectin (MBL), vascular endothelial growth factor (VEGF), human leukocyte antigen (HLA), toll-like receptor (TLR), programmed cell death-1 (PDL-1), interleukin-10 (IL-10RB) and chitotriosidase (CHIT1) increase susceptibility to BF [5,6,7,8,9,10].
Chitin is a polymer of N-Acetyl-d-glucosamine, which is a component of many pathogens, e.g., bacteria, fungi, as well as nematodes. In lymphatic filarial parasites, chitin is a component of microfilaria sheath and important for embryogenesis in adult female worms [11,12]. CHIT1 is a mammalian chitinase which is produced by activated macrophages [13]. CHIT1 plays an important role in innate immunity that degrades chitin-containing pathogens, such as Candida albicans and Madurella mycetomatis, by breaking down the β-1,4-glycosidic bonds of chitin [14,15]. Data suggest that polymorphisms of CHIT1, contributing inactive CHIT protein, might increase susceptibility to lymphatic filarial parasites [16,17]. There are two common polymorphisms of CHIT1 in Asian populations [18], including 24 bp duplication in exon 10 (rs772902435), and p. G102S in exon 4 (rs750476665). The 24 bp duplication in exon 10 causes an abnormal mRNA splicing of CHIT1 and results in the production of inactive truncated CHIT1 [18]. The p. G102S is the substitution of an adenine base (A) on guanine base (G) at nucleotide position 304 of CHIT1 resulting in partially decreasing of CHIT1 efficiency and specificity [18]. It is possible that the CHIT1 deficiency increases the susceptibility to filarial parasites since chitin is a component of microfilaria sheath. However, there are discrepancies of CHIT1 polymorphism. Polymorphism of the 24 bp duplication in exon 10 of CHIT1 increases the susceptibility to BF and filarial chyluria in the Southern and Northern India, respectively [6,19]. However, the 24 bp duplication is not associated with BF in the Papua New Guinea study [20].
Interestingly, there are discrepancies of the association between CHIT1 polymorphisms and susceptibility to BF in different ethnic populations [6,19,20]. The study in Southern India showed that the 24 bp duplication of CHIT1 increases the susceptibility to BF [6] and associated with the filarial chyluria in the Northern India patients [18]. However, the 24 bp duplication of the CHIT1 is not associated with BF in Papua New Guinea [12] In this study, we aimed to investigate whether the 24 bp duplication in exon 10 of CHIT1 and the p. G102S in exon 4 were associated with susceptibility to BF in the Thai–Myanmar border populations.

2. Results

2.1. Demographic and Pathology Information of Study Participants

To investigate the association between CHIT1 polymorphisms (Table 1) and BF. Total 88 individuals in endemic areas of BF, Mae Sot district, Tak province (Thai–Myanmar border), were recruited. The mean age of study participants was not statistically different between cases (29.9) and controls (30.8) (Table 2). 58% of them were males and 42% were females. Males and females were comparable in both BF patients and endemic normal (EN) groups. All participants in both groups had no prior anti-filarial treatment.

2.2. Genotyping of CHIT1 Polymorphisms: 24 bp Duplication and p. G102S

The 24 bp duplication in exon 10 were genotyped by using allele-specific PCR from genomic DNA, extracted from each individuals’ blood, as described in Section 4.2 NA extraction from blood samples. Gel electrophoresis patterns of this polymorphism were shown in Figure 1A. The 24 bp duplication insertion homozygous (INS/INS) showed a single band of 99 bp, whereas wild-type homozygous (WT/WT) represented a single band of 55 bp. The 24 bp duplication heterozygous (WT/INS) represented two bands of 55 and 79 bp.
The p. G102S in exon 4 were genotyped by using PCR sequencing. Chromatograms of DNA sequences were shown in Figure 1B. p. G102S homozygous (A/A) displayed a peak of an adenine base (A) at nucleotide position 304 in the chromatogram, whereas p. G102S wild-type homozygous (G/G) showed a peak of a guanine base (G). The p. G102S heterozygous (G/A) had a double peak of guanine (G) and adenine (A) in the chromatogram (Figure 1B).

2.3. Association between CHIT1 Polymorphisms and Susceptibility to Bancroftian Filariasis in Thai and Myanmar Populations

2.3.1. Genotype Frequencies of CHIT1 Polymorphisms in BF Patients and EN

To determine susceptibility to W. bancrofti infection, genotype and allele frequencies of CHIT1 polymorphisms in EN, and BF patients were studied. Genotype frequencies of CHIT1 polymorphisms, 24 bp duplication and p. G102S, showed no association with BF patients (Table 3).
Unexpectedly, genotype frequencies of 24 bp duplication insertion homozygous (INS/INS) in EN (40.0%) were higher than BF patients (31.4%), but no statistical significance. Genotype frequencies of wild-type homozygous (WT/WT) were overrepresented in BF patients (17.2%) compared with EN (11.1%), while heterozygous (WT/INS) were comparable in both groups (51.4% and 48.9%).
Genotype frequencies of p. G102S homozygous (A/A) in BF patients (21.6%) was higher than in EN (19.0%), without statistical significance (Table 3). Genotype frequencies of p. G102S wild-type homozygous (G/G) was also higher in BF patients (43.1%) than in EN (40.5%), but heterozygous (G/A) was lower in BF patients (35.3%) than in ENs (40.5%), without statistical significance.

2.3.2. Allele Frequencies of CHIT1 Polymorphisms in BF Patients and EN

Allele frequencies of CHIT1 polymorphisms, 24 bp duplication and p. G102S, showed no association with BF patients (Table 4) were studied. Allele frequencies of 24 bp duplication were also higher in EN (64.4%), compared with BF patients (57.1%) (Table 4). The allele frequency of p. G102S in BF patients (39.2%) was similar to EN (39.2%) (Table 4).

3. Discussion

Lymphatic filariasis is a leading cause of permanent disability worldwide. The variability of prevalence and clinical outcomes of lymphatic filariasis are affected by socioeconomic status, personal activities, age, gender and host genetics [2,3,4]. The polymorphisms of host defense pathway genes, including MBL, VEGF, HLA, TLR, PDL-1i and IL-10RB are associated with susceptibility and clinical status of the disease [5,6,7,8,9,10].
CHIT1, produced by activated macrophages, plays a role in degrading chitin-containing pathogens, such as C. albicans and M. mycetomatis, as well as microfilariae of the filarial parasite, W. bancrofti [14]. CHIT1 deficiency, due to the polymorphisms, increases the susceptibility to BF and filarial chyluria in Southern and Northern India, respectively [6,19]. However, the 24 bp duplication is not associated with BF in Papua New Guinea study [20]. Therefore, we investigated the association between BF and CHIT1 polymorphisms, including 24 bp duplication, and p. G102S, to BF in the Thai and Myanmar border populations. These polymorphisms were selected based on association with protein activity and high frequencies in Asian populations.
Our results showed no association between BF and polymorphisms of CHIT1, both 24 bp duplication and p. G102S, consistent with the study in Papua New Guinea. Conversely, these results were not related to the study in South India. These discrepancies may due to several factors. Our infected individuals were recruited based on microfilaria, and circulating filarial antigen, while the Southern India study is a combination of the microfilaria, circulating filarial antigen, and chronic lymphatic disease. Furthermore, the association is not found when these infected groups are analyzed separately. In addition, subjects in the Southern India study had previously been treated with the anti-filarial drug, whereas all subjects in this study had not previously received the anti-filarial treatment.
Genotype frequencies of 24 bp duplication in the Thai and Myanmar populations were also different from both studies in Southern India and Papua New Guinea. While 24 bp duplication homozygous (INS/INS) is rare (<1%) in the Papua New Guinea study (both healthy controls and infected group), it is higher in infected groups in Northern and Southern India, compared with healthy controls [6,19,20]. In contrast to our study, 24 bp duplication homozygous (INS/INS) was overrepresented in EN. The data implicate the importance of genetic epidemiology factor. Allele frequencies of 24 bp duplication of Thai and Myanmar populations were higher than Northern, Southern India and Papua New Guinea populations (0.6125, 0.462, 0.442 and 0.121, respectively). Allele frequencies of 24 bp duplication of Thai and Myanmar populations were similar to healthy individuals in Korea (0.57), Japan (0.54), and Southern China (0.64) [18,22]. Interestingly, the allele frequency of 24 bp duplication is low or absent in Africa, where is an endemic area of lymphatic filariasis [23]. It is possible that other polymorphisms of CHIT1 and other genes might be associated with susceptibility to BF. There is an accordant report concluded that 24 bp duplication does not play a crucial role in protection against hookworm infection, as determined by fecal egg counts [24]. Furthermore, other chitinases and immune responses may play a role against lymphatic filarial parasites.
The discrepancy of genetic polymorphisms of 24 bp duplication in different areas and ethnic groups for other diseases has been reported. The heterozygous genotype of 24 bp duplication is associated with asthma in Northern India [25] but not in the German study [26]. Allele frequencies of 24 bp duplication of Northern India and German are 0.429 and 0.212, respectively. It suggests that different areas and allele frequencies affect the association between the genetic polymorphism and susceptibility to diseases. Moreover, 24 bp duplication is associated with tuberculosis in Europe ancestry, while not in Asian ancestry (0.17 and 0.56, respectively) [18].
In addition, the catalytic efficiency of CHIT1 is different between ethnic groups, due to environmental factors. The catalytic efficiency of wild-type homozygous (WT/WT), and heterozygous (WT/INS) genotype of CHIT1 in Asian populations is significantly lower than Europe and African populations [18]. This data demonstrates that the 24 bp duplication wild-type homozygous (WT/WT) and heterozygous (WT/INS) genotype of CHIT1 are more susceptible to lymphatic filariasis in Thai and Myanmar populations than other populations. This data supports our results, which found overrepresented of wild-type homozygous (WT/WT) in BF patients.
The 24 bp duplication causes an abnormal mRNA splicing of CHIT1 resulting in the production of inactive truncated CHIT1 [18]. However, 24 bp duplication was not associated with susceptibility to BF in our study. We investigated the association between p. G102S of CHIT1 and susceptibility to BF. The p. G102S is a missense mutation that partially decreases efficiency and specificity of CHIT1. Also, the previous report reveals that p. G102S can be found in the Asian population, compared with other polymorphisms (p. G354R, p. 442G and p. A442V). However, this polymorphism was not associated with susceptibility to BF in this study. Additional studies of p. G102S are required in other ethnic populations.
Our results demonstrated the variability of genetic polymorphisms in different ethnic groups. The genetic heterogeneity may result from ethnic, and cultural divisions. This results also suggest that genotyping of CHIT1 polymorphisms, including 24 bp duplication and p. G102S, should not be used to determine the susceptibility of BF in Asia. However, a disadvantage of our study is the number of respondents. Additional studies of CHIT1, and other host defense pathway genes with a higher number of respondents in the Thai and Myanmar populations, and other endemic areas will be required to prove the discrepancy. In addition, it is possible that other chitinase enzyme may play a role against lymphatic filarial parasites. Acidic mammalian chitinase (AMCase) is also able to degrade chitin [27]. It is mainly produced by activated macrophage and acts downstream of IL-13, which is involved in the Th2 response. Previously, AMCase has been demonstrated the protective role against asthma in Caucasian [28]. The chitinase-like proteins, such as YKL-40 and CHI3L1, are also described. They have the ability to bind chitin. Although chitinase-like proteins are not able to degrade chitin [27], chitinase-like proteins are also produced by activated macrophage, suggesting the protective roles in host defense. Further studies to demonstrate the exact mechanism of another chitinase to the lymphatic filarial parasites will be required.

4. Materials and Methods

4.1. Study Areas and Populations

Blood samples were collected from the Thai and Myanmar individuals at Mae Sot district, Tak province (Thai–Myanmar border), endemic areas of BF, were recruited. By using the detection of microfilariae and W. bancrofti specific antigen (NOW® ICT Filariasis Test or the Og4C3 ELISA), a total of 88 individuals was recruited. Characteristics of the study population are shown in Table 2. This study was approved by the Ethics Committee of the Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand (IRB No. 433/50).

4.2. DNA Extraction from Blood Samples

The genomic DNA was extracted from patients’ EDTA blood using modified salting out method. Briefly, EDTA blood samples were resuspended in red blood cell lysis buffer (70 mM NH4Cl and 4 mM Tris-base). Buffy coats were then resuspended in nuclei lysis buffer (400 mM NaCl, 10 mM Tris–HCl and 2 mM EDTA, pH 8.2), and digested with 1% SDS and 1 mg/mL proteinase K at 65 °C for 2 h. Proteins were then removed by using 5.3 M NaCl, following by centrifugation. Finally, genomic DNA was extracted by isopropanol, washed with 70% ethanol, and resuspended in TE buffer.

4.3. Genotyping for CHIT1 Polymorphisms

The 24 bp duplication was genotyped by using allele-specific PCR. Primers flanking the region containing 24 bp duplication were designed, as previously described (Table 1) [6]. PCR reactions were performed with the following conditions: 94 °C for 5 min, 35 cycles of 94 °C for 30 s, 54 °C for 30 s, 72 °C for 30 s, and a final extension at 72 °C for 5 min. PCR products were separated on a 10% polyacrylamide gel using 80 Volt for 1 h, and stained with ethidium bromide. The p. G102S was genotyped by using PCR sequencing (Applied Biosystems 3730xl DNA analyzer). The polymorphic regions containing nucleotide position 304 of the CHIT1 were amplified using the following conditions: 94 °C for 5 min, 35 cycles of 94 °C for 30 s, 60.5 °C for 30 s, 72 °C for 30 s, and a final extension at 72 °C for 5 min. PCR products were subjected to DNA sequencing.

4.4. Statistical Analysis

All statistical analysis was performed using SPSS software for Windows (version 17, IBM, Armonk, NY, USA). A number of chromosomes and subjects were directly counted to determine allele, and genotype frequencies, respectively. Logistic regression and chi-square (χ2) test were used to assess the association between the CHIT1 polymorphisms and susceptibility to BF. A p value of < 0.05 was considered statistically significant.

Author Contributions

Conceptualization, methodology, resources, supervision, writing—review and editing, V.S.; investigation, writing—original draft, S.C.; project administration, supervision, resources, writing—review and editing, funding acquisition, S.N.; all authors read and agreed with the final version of this the paper.

Funding

This research was supported by the Ratchadapiseksompotch funding, Faculty of Medicine, Chulalongkorn University.

Acknowledgments

We thank staffs of Lymphatic Filariasis and Tropical Medicine Research Unit, Faculty of Medicine, Chulalongkorn University, for collecting field samples.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Nuchprayoon, S. DNA-based diagnosis of lymphatic filariasis. Southeast Asian J. Trop. Med. Public Health 2009, 40, 904–913. [Google Scholar]
  2. Das, P.K.; Manoharan, A.; Srividya, A.; Grenfell, B.T.; Bundy, D.A.; Vanamail, P. Frequency distribution of Wuchereria bancrofti microfilariae in human populations and its relationships with age and sex. Parasitology 1990, 101 Pt 3, 429–434. [Google Scholar] [CrossRef]
  3. Subrahmanyam, D.; Mehta, K.; Nelson, D.S.; Rao, Y.V.; Rao, C.K. Immune reactions in human filariasis. J. Clin. Microbiol. 1978, 8, 228–232. [Google Scholar]
  4. Tisch, D.J.; Hazlett, F.E.; Kastens, W.; Alpers, M.P.; Bockarie, M.J.; Kazura, J.W. Ecologic and biologic determinants of filarial antigenemia in bancroftian filariasis in Papua New Guinea. J. Infect. Dis. 2001, 184, 898–904. [Google Scholar] [CrossRef]
  5. Chan, S.H.; Dissanayake, S.; Mak, J.W.; Ismail, M.M.; Wee, G.B.; Srinivasan, N.; Soo, B.H.; Zaman, V. HLA and filariasis in Sri Lankans and Indians. Southeast Asian J. Trop. Med. Public Health 1984, 15, 281–286. [Google Scholar]
  6. Choi, E.H.; Zimmerman, P.A.; Foster, C.B.; Zhu, S.; Kumaraswami, V.; Nutman, T.B.; Chanock, S.J. Genetic polymorphisms in molecules of innate immunity and susceptibility to infection with Wuchereria bancrofti in South India. Genes Immun. 2001, 2, 248–253. [Google Scholar] [CrossRef] [PubMed]
  7. Junpee, A.; Tencomnao, T.; Sanprasert, V.; Nuchprayoon, S. Association between Toll-like receptor 2 (TLR2) polymorphisms and asymptomatic bancroftian filariasis. Parasitol. Res. 2010, 107, 807–816. [Google Scholar] [CrossRef] [PubMed]
  8. Meyrowitsch, D.W.; Simonsen, P.E.; Garred, P.; Dalgaard, M.; Magesa, S.M.; Alifrangis, M. Association between mannose-binding lectin polymorphisms and Wuchereria bancrofti infection in two communities in North-Eastern Tanzania. Am. J. Trop. Med. Hyg. 2010, 82, 115–120. [Google Scholar] [CrossRef] [PubMed]
  9. Panda, A.K.; Sahoo, P.K.; Kerketta, A.S.; Kar, S.K.; Ravindran, B.; Satapathy, A.K. Human lymphatic filariasis: Genetic polymorphism of endothelin-1 and tumor necrosis factor receptor II correlates with development of chronic disease. J. Infect. Dis. 2011, 204, 315–322. [Google Scholar] [CrossRef] [PubMed]
  10. Venugopal, G.; O’Regan, N.L.; Babu, S.; Schumann, R.R.; Srikantam, A.; Merle, R.; Hartmann, S.; Steinfelder, S. Association of a PD-L2 Gene Polymorphism with Chronic Lymphatic Filariasis in a South Indian Cohort. Am. J. Trop. Med. Hyg. 2018. [Google Scholar] [CrossRef] [PubMed]
  11. Harris, M.T.; Lai, K.; Arnold, K.; Martinez, H.F.; Specht, C.A.; Fuhrman, J.A. Chitin synthase in the filarial parasite, Brugia malayi. Mol. Biochem. Parasitol. 2000, 111, 351–362. [Google Scholar] [CrossRef]
  12. Fuhrman, J.A.; Piessens, W.F. Chitin synthesis and sheath morphogenesis in Brugia malayi microfilariae. Mol. Biochem. Parasitol. 1985, 17, 93–104. [Google Scholar] [CrossRef]
  13. Di Rosa, M.; Malaguarnera, G.; De Gregorio, C.; Drago, F.; Malaguarnera, L. Evaluation of CHI3L-1 and CHIT-1 expression in differentiated and polarized macrophages. Inflammation 2013, 36, 482–492. [Google Scholar] [CrossRef]
  14. Kanneganti, M.; Kamba, A.; Mizoguchi, E. Role of chitotriosidase (chitinase 1) under normal and disease conditions. J. Epithel. Biol. Pharmacol. 2012, 5, 1–9. [Google Scholar] [CrossRef]
  15. Labadaridis, I.; Dimitriou, E.; Theodorakis, M.; Kafalidis, G.; Velegraki, A.; Michelakakis, H. Chitotriosidase in neonates with fungal and bacterial infections. Arch. Dis. Child. Fetal Neonatal Ed. 2005, 90, F531–F532. [Google Scholar] [CrossRef]
  16. Lehrnbecher, T.; Bernig, T.; Hanisch, M.; Koehl, U.; Behl, M.; Reinhardt, D.; Creutzig, U.; Klingebiel, T.; Chanock, S.J.; Schwabe, D. Common genetic variants in the interleukin-6 and chitotriosidase genes are associated with the risk for serious infection in children undergoing therapy for acute myeloid leukemia. Leukemia 2005, 19, 1745–1750. [Google Scholar] [CrossRef]
  17. Verwer, P.E.; Notenboom, C.C.; Eadie, K.; Fahal, A.H.; Verbrugh, H.A.; van de Sande, W.W. A Polymorphism in the Chitotriosidase Gene Associated with Risk of Mycetoma Due to Madurella mycetomatis Mycetoma--A Retrospective Study. PLoS Negl. Trop. Dis. 2015, 9, e0004061. [Google Scholar] [CrossRef] [PubMed]
  18. Lee, P.; Waalen, J.; Crain, K.; Smargon, A.; Beutler, E. Human chitotriosidase polymorphisms G354R and A442V associated with reduced enzyme activity. Blood Cells Mol. Dis. 2007, 39, 353–360. [Google Scholar] [CrossRef] [PubMed]
  19. Pant, S.; Agarwal, J.; Goel, A.; Gangwar, P.K.; Waseem, M.; Gupta, P.; Sankhwar, S.N.; Purkait, B. Does 24bp Duplication of Human CHIT1 Gene (Chitotriosidase1) Predispose to Filarial Chyluria? A Case-Control Study. J. Clin. Diagn. Res. 2016, 10, PC01–PC04. [Google Scholar] [CrossRef] [PubMed]
  20. Hise, A.G.; Hazlett, F.E.; Bockarie, M.J.; Zimmerman, P.A.; Tisch, D.J.; Kazura, J.W. Polymorphisms of innate immunity genes and susceptibility to lymphatic filariasis. Genes Immun. 2003, 4, 524–527. [Google Scholar] [CrossRef]
  21. Boot, R.G.; Renkema, G.H.; Verhoek, M.; Strijland, A.; Bliek, J.; de Meulemeester, T.M.; Mannens, M.M.; Aerts, J.M. The human chitotriosidase gene. Nature of inherited enzyme deficiency. J. Biol. Chem. 1998, 273, 25680–25685. [Google Scholar] [CrossRef]
  22. Woo, K.H.; Lee, B.H.; Heo, S.H.; Kim, J.M.; Kim, G.H.; Kim, Y.M.; Kim, J.H.; Choi, I.H.; Yang, S.H.; Yoo, H.W. Allele frequency of a 24 bp duplication in exon 10 of the CHIT1 gene in the general Korean population and in Korean patients with Gaucher disease. J. Hum. Genet. 2014, 59, 276–279. [Google Scholar] [CrossRef] [PubMed]
  23. Arndt, S.; Hobbs, A.; Sinclaire, I.; Lane, A.B. Chitotriosidase deficiency: A mutation update in an african population. JIMD Rep. 2013, 10, 11–16. [Google Scholar]
  24. Hall, A.J.; Quinnell, R.J.; Raiko, A.; Lagog, M.; Siba, P.; Morroll, S.; Falcone, F.H. Chitotriosidase deficiency is not associated with human hookworm infection in a Papua New Guinean population. Infect. Genet. Evol. 2007, 7, 743–747. [Google Scholar] [CrossRef] [PubMed]
  25. Sinha, S.; Singh, J.; Jindal, S.K.; Birbian, N.; Singla, N. Association of 24 bp duplication of human CHIT1 gene with asthma in a heterozygous population of north India: A case-control study. Lung 2014, 192, 685–691. [Google Scholar] [CrossRef] [PubMed]
  26. Bierbaum, S.; Superti-Furga, A.; Heinzmann, A. Genetic polymorphisms of chitotriosidase in Caucasian children with bronchial Asthma. Int. J. Immunogenet. 2006, 33, 201–204. [Google Scholar] [CrossRef] [PubMed]
  27. Malaguarnera, L. Chitotriosidase: The yin and yang. Cell Mol. Life Sci. 2006, 63, 3018–3029. [Google Scholar] [CrossRef]
  28. Bierbaum, S.; Nickel, R.; Koch, A.; Lau, S.; Deichmann, K.A.; Wahn, U.; Superti-Furga, A.; Heinzmann, A. Polymorphisms and haplotypes of acid mammalian chitinase are associated with bronchial asthma. Am. J. Respir. Crit. Care Med. 2005, 172, 1505–1509. [Google Scholar] [CrossRef]
Pathogens 08 00041 g001 550
Figure 1. Genotyping of CHIT1 polymorphisms. (A) 10% polyacrylamide gel electrophoresis patterns of 24 bp duplication in exon 10. PCR products were separated with 10% polyacrylamide gel (in TAE buffer) at 80 Volt for 1 h. Lane 1: 50 bp DNA ladder (M); lane 2: wild-type homozygous (WT/WT); lane 3: mutant homozygous (INS/INS); and lane 4: heterozygous (WT/INS) (B) Chromatogram of DNA sequences of p. G102S in exon 4. Top: wild-type homozygous (G/G); Middle: heterozygous (G/A); Bottom: mutant homozygous (A/A).
Figure 1. Genotyping of CHIT1 polymorphisms. (A) 10% polyacrylamide gel electrophoresis patterns of 24 bp duplication in exon 10. PCR products were separated with 10% polyacrylamide gel (in TAE buffer) at 80 Volt for 1 h. Lane 1: 50 bp DNA ladder (M); lane 2: wild-type homozygous (WT/WT); lane 3: mutant homozygous (INS/INS); and lane 4: heterozygous (WT/INS) (B) Chromatogram of DNA sequences of p. G102S in exon 4. Top: wild-type homozygous (G/G); Middle: heterozygous (G/A); Bottom: mutant homozygous (A/A).
Pathogens 08 00041 g001
Table
Table 1. Selected single nucleotide polymorphisms of CHIT1 and primer sequences for genotyping.
Table 1. Selected single nucleotide polymorphisms of CHIT1 and primer sequences for genotyping.
PolymorphismSNP IDSNP InfluencePrimer Sequences
24 bp duplicationRs
772902435		Associated with an abnormal mRNA splicing of CHIT1 and results in the production of inactive truncated CHIT1 [21]F 5′-AGC TAT CTG AAG CAG AAG-3′
R 5′-GGA GGA GCC GGC AAA GTC-3′ [6]
p. G102SRs
750476665		Associated with the substitution of glycine (Gly) to serine (Ser) at the amino acid position 102 of CHIT1, resulting in decreasing of CHIT1 enzymatic activity and specificity [21]F 5′-GGC AGC TGG CAG AGT AAA TCC-3′
R 5′-CCC AGA AGG AAA TTC AGC CC-3′
Table
Table 2. Characteristics of total of 88 individuals from the Thai and Myanmar individuals at Western Thailand.
Table 2. Characteristics of total of 88 individuals from the Thai and Myanmar individuals at Western Thailand.
CharacteristicsBancroftian Filariasis PatientsEndemic Normal
Gender (M/F)27/2419/18
Median age (range)30.6 (16–61)31.8 (18–68)
Mean age (±SD)29.9 ± 12.530.8 ± 12.9
TreatmentNoNo
Bed net usageYesYes
PathologyNoneNone
Ag Test (ICT card test and Og4C3 ELISA)All positiveAll negative
W. bancrofti circulating antigen levels (U/mL, range)29,986 (133–71,339)<32 (<32)
Table
Table 3. Genotype frequencies of 24 bp duplication in exon 10 and p. G102S in exon 4 polymorphisms of CHIT1 in BF patients and EN.
Table 3. Genotype frequencies of 24 bp duplication in exon 10 and p. G102S in exon 4 polymorphisms of CHIT1 in BF patients and EN.
GenotypeNumber of Endemic Normal (%)Number of Bancroftian Filariasis Patients (%)p Value
24 bp duplicationWT/WT5 (11.1%)6 (17.2%)0.623
WT/INS22 (48.9%)18 (51.4%)
INS/INS18 (40.0%)11 (31.4%)
Total45 (100.0%)35 (100.0%)
p. G102SG/G15 (40.5%)22 (43.1%)0.876
G/A15 (40.5%)18 (35.3%)
A/A7 (19.0%)11 (21.6%)
Total37 (100.0%)51 (100.0%)
WT/WT = wild-type homozygous, WT/INS = 24 bp duplication heterozygous. INS/INS = 24 bp mutant homozygous, G/G = p. G102S wild-type homozygous, G/A = p. G102S heterozygous, A/A = p. G102S homozygous.
Table
Table 4. Allele frequencies of 24 bp duplication in exon 10 and p. G102S in exon 4 polymorphisms of CHIT1 in BF patients and EN.
Table 4. Allele frequencies of 24 bp duplication in exon 10 and p. G102S in exon 4 polymorphisms of CHIT1 in BF patients and EN.
GenotypeNumber of Endemic Normal (%)Number of Bancroftian Filariasis Patients (%)p Value
24 bp duplicationWT32 (35.6%)30 (42.9%)0.347
INS58 (64.4%)40 (57.1%)
Total90 (100.0%)70 (100.0%)
p. G102SG45 (60.8%)62 (60.8%)0.997
A29 (39.2%)40 (39.2%)
Total74 (100.0%)102 (100.0%)
WT = wild-type allele of 24 bp duplication in exon 10 of CHIT1, INS = mutant allele of 24 bp duplication in exon 10 of CHIT1, G = wild-type allele of p. G102S in exon 4 of CHIT1, A = mutant allele of p. G102S in exon 4 of CHIT1.

Share and Cite

MDPI and ACS Style

Sanprasert, V.; Charuchaibovorn, S.; Nuchprayoon, S. The Genetic Polymorphisms of 24 Base Pair Duplication and Point G102S of Human Chitotriosidase to Bancroftian Filariasis at the Thai–Myanmar Border. Pathogens 2019, 8, 41. https://doi.org/10.3390/pathogens8010041

AMA Style

Sanprasert V, Charuchaibovorn S, Nuchprayoon S. The Genetic Polymorphisms of 24 Base Pair Duplication and Point G102S of Human Chitotriosidase to Bancroftian Filariasis at the Thai–Myanmar Border. Pathogens. 2019; 8(1):41. https://doi.org/10.3390/pathogens8010041

Chicago/Turabian Style

Sanprasert, Vivornpun, Sarit Charuchaibovorn, and Surang Nuchprayoon. 2019. "The Genetic Polymorphisms of 24 Base Pair Duplication and Point G102S of Human Chitotriosidase to Bancroftian Filariasis at the Thai–Myanmar Border" Pathogens 8, no. 1: 41. https://doi.org/10.3390/pathogens8010041

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop