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Journal : UNEJ e-Proceeding

Imunogenic Protein of Salivary Gland from Anopheles sundaicus Armiyanti, Yunita; Nuryady, Moh Mirza; Utomo, Sugeng Setyo; Sardjono, Teguh Wahju; Fitri, Loeki Enggar; Senjarini, Kartika
UNEJ e-Proceeding Indonesian Protein Society (IPS), International Seminar and Workshop 2014
Publisher : UNEJ e-Proceeding

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Abstract

Malaria is still a major problem for developing countries, including Indonesia. One approach to overcome this disease is prevention by vaccination. However, there is still no effective malaria vaccine that is applicable. The ideal malaria vaccine is a combination vaccine that can prevent the pre-erythrocytic cycle, the erythrocytic cycle and transmission process. Salivary vector-based vaccine has the potential to be developed as a malaria vaccine because it can prevent transmission process and also decrease the morbidity of the disease. Saliva from Anopheles contains vasomodulator and immunomodulatory components, that are required in the blood feeding process, but in the same time it could enhance the transmission of the malaria parasite. If the component in the salivary vector can increase pathogen infection, then vaccinating the host with its anti-substances can control the transmission of pathogens (Transmision Blocking Vaccine). Anopheles sundaicus is an important vector of malaria in coastal areas of Java, Bali, Sumatra, Kalimantan and West Nusa Tenggara islands. Repeated exposures of Salivary Gland Extract (SGE) from this vector have been proven to be able to decrease parasitemic rates in mouse model for malaria in our study. The objective of this research is to determine and localize the immunogenic protein from SGE of An. sundaicus as the first step for the characterization of its immunomodulatory component. Mosquito salivary gland protein profile of An.sundaicus was determine by SDS-PAGE. Determination of salivary glands immunogenic proteins was conducted by Western Blotting with IgG from people living from endemic area as primary antibody. Out of 15 bands appeared in SDS PAGE ranging from 24 kD to 138 kD, only two protein bands with  molecular weights of 68 and 37 kDa were the most immunogenic. Those immunogenic proteins were consistent recognized by pooled serum of people as well as by individual response. Keywords: malaria, saliva, vector, immunogenic protein, vaccine
Specific sequence of Plasmodium falciparum DBL domains associated with severe malaria outcome Sulistyaningsih, Erma; Fitri, Loeki Enggar; Loescher, Thomas; Berens-Riha, Nicole
UNEJ e-Proceeding Indonesian Protein Society (IPS), International Seminar and Workshop 2014
Publisher : UNEJ e-Proceeding

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Abstract

Duffy-binding like (DBL) domains of Plasmodium falciparum are believed to be involved in erythrocytes invasion and infected erythrocytes cytoadhesion during the blood stage of malaria infection. In Plasmodium falciparum, DBL domains found in the two different protein families; Erythrocyte Binding Ligand (EBL) including EBA-175, EBA-140, EBA-181 and EBL-1, and Plasmodium falciparum Erythrocyte Membrane Protein-1. The study aimed at investigating the specific sequence of Plasmodium falciparum involved in severe malaria outcome.Blood samples from severe and uncomplicated falciparum malaria cases from Papua and South Kalimantan province, Indonesia, were collected for DNA extraction. A dried blood on filter paper were used for RNA extraction. PCR was performed using UNIEBP primers and directly sequenced. Internal var D primers were designed according to the sequencing of the ~550 bp band produced by UNIEBP primers. The nucleotide sequences were analyzed by NCBI BLAST. Multiple bands ranging from nearly 250 bp to 1 kb were resulted from gDNA in all samples. Two isolates yielded bands of 450 and 525 bp, three isolates showed three bands additionally 250 bp, one isolate presented four bands additionally 800 bp and one isolate resulted one band additionally 1 kb.Amplification of cDNA from severe malaria cases produced one to four bands ranging from 250 bp to 700 bp, and no band observed from cDNA of uncomplicated malaria. Sequencing of the 418 bp bands matched with the eba-175 gene, the 316 bp determined as DBL1a domain and 486 bp band matched with the DBLg domain isolated from placenta of PAM’s Malawian woman. The expression of a 237 bp sequence corresponding to var D gene, was detected solely in severe malaria patients, implicating an association of gene expression and manifestation of severe malaria. Further characterization of the var D gene with a larger sample size is required to draw a definite conclusion. Keywords: DBL domains, Plasmodium falciparum, severe malaria, var D gene.
Imunogenic Protein of Salivary Gland from Anopheles sundaicus Yunita Armiyanti; Moh Mirza Nuryady; Sugeng Setyo Utomo; Teguh Wahju Sardjono; Loeki Enggar Fitri; Kartika Senjarini
UNEJ e-Proceeding Indonesian Protein Society (IPS), International Seminar and Workshop 2014
Publisher : UPT Penerbitan Universitas Jember

Show Abstract | Download Original | Original Source | Check in Google Scholar

Abstract

Malaria is still a major problem for developing countries, including Indonesia. One approach to overcome this disease is prevention by vaccination. However, there is still no effective malaria vaccine that is applicable. The ideal malaria vaccine is a combination vaccine that can prevent the pre-erythrocytic cycle, the erythrocytic cycle and transmission process. Salivary vector-based vaccine has the potential to be developed as a malaria vaccine because it can prevent transmission process and also decrease the morbidity of the disease. Saliva from Anopheles contains vasomodulator and immunomodulatory components, that are required in the blood feeding process, but in the same time it could enhance the transmission of the malaria parasite. If the component in the salivary vector can increase pathogen infection, then vaccinating the host with its anti-substances can control the transmission of pathogens (Transmision Blocking Vaccine). Anopheles sundaicus is an important vector of malaria in coastal areas of Java, Bali, Sumatra, Kalimantan and West Nusa Tenggara islands. Repeated exposures of Salivary Gland Extract (SGE) from this vector have been proven to be able to decrease parasitemic rates in mouse model for malaria in our study. The objective of this research is to determine and localize the immunogenic protein from SGE of An. sundaicus as the first step for the characterization of its immunomodulatory component. Mosquito salivary gland protein profile of An.sundaicus was determine by SDS-PAGE. Determination of salivary glands immunogenic proteins was conducted by Western Blotting with IgG from people living from endemic area as primary antibody. Out of 15 bands appeared in SDS PAGE ranging from 24 kD to 138 kD, only two protein bands with  molecular weights of 68 and 37 kDa were the most immunogenic. Those immunogenic proteins were consistent recognized by pooled serum of people as well as by individual response. Keywords: malaria, saliva, vector, immunogenic protein, vaccine
Specific sequence of Plasmodium falciparum DBL domains associated with severe malaria outcome Erma Sulistyaningsih; Loeki Enggar Fitri; Thomas Loescher; Nicole Berens-Riha
UNEJ e-Proceeding Indonesian Protein Society (IPS), International Seminar and Workshop 2014
Publisher : UPT Penerbitan Universitas Jember

Show Abstract | Download Original | Original Source | Check in Google Scholar

Abstract

Duffy-binding like (DBL) domains of Plasmodium falciparum are believed to be involved in erythrocytes invasion and infected erythrocytes cytoadhesion during the blood stage of malaria infection. In Plasmodium falciparum, DBL domains found in the two different protein families; Erythrocyte Binding Ligand (EBL) including EBA-175, EBA-140, EBA-181 and EBL-1, and Plasmodium falciparum Erythrocyte Membrane Protein-1. The study aimed at investigating the specific sequence of Plasmodium falciparum involved in severe malaria outcome.Blood samples from severe and uncomplicated falciparum malaria cases from Papua and South Kalimantan province, Indonesia, were collected for DNA extraction. A dried blood on filter paper were used for RNA extraction. PCR was performed using UNIEBP primers and directly sequenced. Internal var D primers were designed according to the sequencing of the ~550 bp band produced by UNIEBP primers. The nucleotide sequences were analyzed by NCBI BLAST. Multiple bands ranging from nearly 250 bp to 1 kb were resulted from gDNA in all samples. Two isolates yielded bands of 450 and 525 bp, three isolates showed three bands additionally 250 bp, one isolate presented four bands additionally 800 bp and one isolate resulted one band additionally 1 kb.Amplification of cDNA from severe malaria cases produced one to four bands ranging from 250 bp to 700 bp, and no band observed from cDNA of uncomplicated malaria. Sequencing of the 418 bp bands matched with the eba-175 gene, the 316 bp determined as DBL1a domain and 486 bp band matched with the DBLg domain isolated from placenta of PAM’s Malawian woman. The expression of a 237 bp sequence corresponding to var D gene, was detected solely in severe malaria patients, implicating an association of gene expression and manifestation of severe malaria. Further characterization of the var D gene with a larger sample size is required to draw a definite conclusion. Keywords: DBL domains, Plasmodium falciparum, severe malaria, var D gene.
Co-Authors A.A. Ketut Agung Cahyawan W AA Sudharmawan, AA Adilah Ulfiati Adilah Ulfiati, Adilah Agustin Iskandar Agustina Tri Endharti Agustina Tri Endharti Ahmad Fauziansyah Ahmad Fauziansyah Zian Ajeng Maharani Putri Alfian Wika Cahyono Ali Rahmanto Anak Agung Gede Sugianthara Andi Arahmadani Arasy Anik Widijanti Arasy, Andi Arahmadani Ardhian Wardana Ardhian Wardana, Ardhian Ariel, Dio Giovanni Arif Setyawan, Sang Aji Arifin, Mochammad Samsul Astutik Pudjirahaju Bagus Hermansyah BR. Sijabat, Melda Fio Flora Bramantyo Aji Wicaksono Bramantyo Aji Wicaksono, Bramantyo Aji Brigitta Ida RV Corebima Burhan, Niniek Cahyono, Alfian Wika Chairul A. Nidom Chairul A. Nidom Dalhar, Mochamad Damayanti, Ria Dara Dasawulansari Syamsuri Desy Andari Dewi Indiastari Didi Candradikusuma Didi Candradikusuma Dio Giovanni Ariel Dorta Simamora Edi Widjajanto Elviliana, Elviliana Erma Sulistyaningsih Erwan, Nabila Erina Eviana Norahmawati Firdiani Nur Afifah, Firdiani Nur Fitria Febriliani Putri Fitriana Nugraheni Fitriana Nugraheni, Fitriana Ghofar, Hervin Febrina Handono Kalim Hayu Sukowati Nopitasari Heri Kristianto Heri Suroso, Heri Herpan Syafii Harahap Hervin Febrina Ghofar Hidayat Sujuti Husnul Asariati Husnul Asariati I Ketut Gede Muliartha Ihwan Ihwan Ika Setyo Rini Irene Ratridewi Isnadiyah Juhdi Joko Agus Gunawan Josef Sem Berth Tuda Juhdi, Isnadiyah Kana Mardhiyyah Kartika Senjarini Karyono Mintaroem Karyono Mintaroem Karyono Mintaroem Khadafi Indrawan Khairiyadi Khairiyadi Krisni Subandiyah Krisni Subandiyah Krisni Subandiyah Krsni Subandiyah Kusworini Handono Laksmi Karunia Tanuwijaya Laksmi Karunia Tanuwijaya Lilik Maslachah Lilik Zuhriyah M. Fahrul Arifin Mardhiyyah, Kana Mario B. Nara Melda Fio Flora BR. Sijabat Merici, Angela Mochammad Dalhar Moh Mirza Nuryady Moh Mirza Nuryady, Moh Mirza Mohammad Saifur Rahman Mulyohadi Ali Muti'ah, Roihatul Nabila Erina Erwan Nafi’ah, Riris Waladatun Namira, Anggi Alya Nashi Widodo Natalia Erica Jahja Nicole Berens-Riha Nicole Berens-Riha, Nicole Niniek Budiarti B Niniek Burhan Noer Aini Nopitasari, Hayu Sukowati Nugraha, Rivo Yudhinata Brian Nur Fahma Pradiptasari Nur Fahma Pradiptasari, Nur Fahma Nur Hidayat Nur Permatasari Nurdiana Nurdiana Nurdiana Nurdiana Nurtyas, Yunika pertiwi, resti anggun Poedji Hastutiek Pratama, Andhika Putra Agus Putri, Ajeng Maharani Putri, Fitria Febriliani Putri, Rachmania Rachmania Putri Rahma, Zainabur Rahmad Rahmad Rahman, Mohammad Saifur Renny Suwarniaty Renny Suwarniaty, Renny Rifa'i, Muhaimin Rivo Yudhinata Brian Nugraha Rizky Amalia Rohma, Novita Ainur Sabrina Sunyoto, Nimas Mayang Saiful Akhyar Lubis Sakunda Anggarini, Sakunda Samsul Arifin Siti Candra Windu Baktiyani Siti Candra Windu Baktiyani Soebaktiningsih Soebaktiningsih Soemarko Soemarko Sri Hayati, Yati Sri Murwani Sri Poeranto, Sri Sri Suhartini Sri Winarsih Subekti, Indah Fitriana Sudjari Sudjari Sugeng Setyo Utomo Sugeng Setyo Utomo, Sugeng Setyo Sujarot Dwi Sasmito sujarot dwi sasmito, sujarot dwi Sumarno . Sumarno Sumarno Susanto Nugroho Susanto Nugroho Sutiman B. Sumitro Swastomo, Rahadi Syarkiah Syarkiah Teguh Wahju Sardjono Teguh Wahju Sardjono Teguh Wahju Sardjono Thomas Loescher Thomas Loescher, Thomas Tikto Wahyono Tinny Endang Endang H Tinny Endang Hernowati Tinny Endang Hernowati Tuda, Josef Sem Berth Umar Zein Verry Asfirizal Vonny Mariany Deckert Wahyono, Tikto Wongso Suhendro Yoes Prijatna Dachlan Yuanita Mulyastuti Yuliyanik yuliyanik Yunika Nurtyas Yunita Armiyanti Zainabur Rahma Zakiyah Zulaifa Zakiyah Zulaifa, Zakiyah