Technology develops rapidly in recent years and it affects on the research diversity and implementation to gain the value of research outcomes. We may realize without the advanced technology, people in the world would be suffered by hunger. However, at the same time researchers also realize that everything resulting from the laboratory should be safe for human and their environment. Therefore, ethical research as a form of researcher’s responsibility should arise in every research work. The purpose of this article is to report the recombinant protein research in Jembrana disease virus (1), to identify the relevant of ethical features to the recombinant protein research as vaccine development (2) and to discuss the integration of utilitarianism principle to vaccine development (3). Jembrana disease virus (JDV) is a fatal and an infectious disease caused by virus that classified as retrovirus from Retrovidae family. The disease called as Jembrana disease attacks specifically in a local beef cattle in Indonesia, namely is Bali cattle (Bos javanicus). The fatal rate of disease caused death of 20% population of bali cattle in the past 1964, and up to present prevention of the disease was performed by vaccination. The jembarana vaccine is used until now is prepared from organs of infected Bali cattle to obtain crude vaccine. Improved vaccine of Jembrana disease is attempted to increase the availability and quality of Jembrana vaccine by implementation of recombinant DNA technology to produce recombinant protein as Jembrana vaccine. This article was prepared from our research laboratory and some examples of related research work from other reports. Bioethics issue is as much as involved to furnish this article as a reminder to scientists in doing their research work. It suggests that this article could be as a contribution of laboratory research to bioethics issue which output from the laboratory works should be safe and useful for live organism and environment.

In the past of 17th century, Aristotle has focused on scientific concerns, and the science has progressed rapidly and wider in molecular biology. Consequences, there are now several societies emerging concomitant with the rapid progress of advanced science in some specific science interests. At the beginning of 21st century, the progress of science and sustainable development of societies are very dependent of each other (Airaksinen, 1995). It was discoursed in the Societas Ethica meeting of science and development progress of societies have warned us that ethics and science has become important element for both interests of science and the interests of society. As reminded by Behm (2005) that research ethics is recognized widely as an important feature of scientific practice. There are three human interests towards knowledge, those are technical, practical and emancipatory (Habermas, 1971). Technical interests can be found behind natural sciences and behind some systematic social sciences, practical interest lies behind humanistic sciences and emancipatory interest lies behind critical social sciences. Based on the Haberman’s interests, it is clear that researchers need to perform their research works wisely and responsible for live organism and environment.

When researchers deal with their research by using live organism or something live then they need to be concern with bioethics. There are many definitions of bioethics, most of them controlling the effect of research works to human and environment. However, when we line up with focus on research purposes then we get the same red line that we need to aware to the environment in wider meaning.

Have you been thinking that someday life in the world could be extinct due to there is no any medicines for recovering or healing of illness. In some sides, the existing of advanced technology based on molecular level offering eases, efficiency and cheaper in mass production. In other sides, there is still a gap of fear resulting from breakthrough of the advanced technology. Mankind have been given mind and soul, in another hand, they also need to improve medical material with higher quality, cheaper, safe and useful for a wider mankind.

Starting from the above statement, bioethics comes in the middle of a higher life demand that have to be fulfilled for sustainability of their life. Therefore, the definition of bioethics needs to be understood carefully and wisely. Bioethics is a combine of several disciplines of knowledge concerning on ethics, legality and social questions (worries) that emerged as the effect of development in medicines, science and biotechnology (Mepham, 2005). Advances in science with molecular-based technology as a tool, this will be useful to cope with the lack of food and medicine availability, as long as the advanced technology and material research are chosen wisely. This option of doing research meets to the definition of bioethics which is stated as the study of moral and social implications of techniques resulting from advances in the biological sciences (Mepham, 2005).

As stated by first bioethicist of Aldo Leopold that “A thing is right when it tends to preserve the integrity, stability, and beauty of the biotic community. It is wrong when it tends otherwise" (Leopold, 1949). This statement reminds us that mankind should be responsible in using the natural sources wisely. Another definition stated that bioethics is actually not a discipline, but it has become a meeting ground for a number of disciplines, discourses, and organizations concerned with ethical, legal, and social questions raised by advances in medicines, science, and biotechnology (O’Neill, 2002; Mepham, 2005). Those definitions are clear that both concerned with implication of using of advanced technology for producing huge number of medical products.

Creating of biological medicines is also needed for animal as another organism in earth, which will be discussed in this article as study case in producing vaccine. Entering biotechnology era, it is very possible to develop the Jembrana vaccine by approaching of a recombinant DNA technology to produce recombinant viral protein as a recombinant vaccine for controlling the jembrana disease in Bali cattle.

Bali cattle is as a local beef cattle in Indonesia and as one of gene pools beef cattle in south-east Asia, it has superiority in reproduction of one year in calving interval. In spite of that reproductive superiority, Bali cattle has another superiority in meat characteristics, i.e., lean, juicy and smooth texture of muscular fiber. However, Bali cattle has inferiority in susceptibility of Jembrana disease virus (JDV), Chadwick et al. (1995). The fatal death rate of Jembrana disease was about 20% of Bali cattle population in 1964 (Moll, 1998). In the past colonization by Dutch, Bali cattle was restricted rearing in Bali island to prevent their genetic purity. By the time and up to present, distribution of Bali cattle is wider throughout of Indonesia islands. Consequence, the disease is transmitted to another islands and the need of Jembrana vaccine is increased. Up to present, vaccination is performed by using crude tissue-derived vaccine (Hartaningsih and Wilcox, 1996; Hartaningsih et al., 2000). Processing of the crude vaccine is costly, due to using infected lymph or spleen organs of fresh slaughtered Bali cattle. Disadvantageous of the crude vaccine is a lower quality, expensive, lower production, demanding in handling to the field.

Advancing in recombinant DNA and more popular technology for medicine research is recombinant protein. Jembrana disease virus (JDV) belongs to lentivirus group of Retroviridae family (Wilcox et al, 1995). The JDV genome (7732bp) consists at least of three major genes (gag, pol and env) encoding proteins that needed for virus replications (Chacdwick et al, 1995). Moreover, it was stated that the env gene encodes TM and SU proteins on the surface of the virus and most attempts to induce a protective immunity against lentivirus infectious. Another immunogenic viral protein is Tat that coded by tat gene. The tat gene is one of small accessory genes and lies between gag and pol genes. In recent study, those three Jembarana genes were cloned into a plasmid vector in a pGEX system. The obtained clones were transformed through a host cell of Escher coli ichia(E. coli) to express their recombinant proteins. The host cell of E. coli is very common in medicine production since they can multiply quicker in cell numbers bearing the recombinant proteins. Therefore, most recombinant protein is always purified from E. coli after cloning on the gene into expression vectors (Studier and Moffatt, 1986).

The study case of recombinant protein research in Jembrana disease virus (JDV)
At least three proteins of SU (superficial unit), TM (transmembrane) and Tat are potential candidates for a protective vaccine against Jembrana disease in Bali cattle (ACIAR report, 2004). The TM and SU proteins are encoded by env gene while Tat protein was encoded by tat gene. By polymerase chair reaction (PCR), those three gene fragments were amplified then constructed in pGEX system. To express proteins therefore the constructs need to be transformed into a host cell of BL21 strain of Escherichia coli as a manufacture of producing the proteins. Production of the recombinant proteins for JDV is possible as the genes encoding for the proteins have been well-characterized (Chadwick et al., 1995).

When cloning of JSU, JTM and JTat were performed using pGEX system in which the proteins were fused with GST tag, the expression of JSU, JTM and JTat protein were also conducted in pGEX system. The molecular size of GST tag is ± 26kDa (Amershampharmacia biotech, 1997), while molecular weights of JSU, JTM and JTat are ±34, ±10 and ±10.7kDa, respectively. Therefore, when proteins is fused with the GST tag, then resulted recombinant proteins of ± 60, ± 36 and ± 36.7kDa respectively for JSU, JTM and JTat. The purification and characterization of those proteins were conducted by SDS-PAGE and Western blotting (WB). Constructing of JTat was also conducted in pET system where the system used a 6-histidine tag which has a molecular size less than 1 kDa. This construction of JTat in pET system was aimed to increase the efficacy of the resulted recombinant protein of JTat. Expression of JTat was transformed in BL21 E. coli.

Those recombinant proteins are the virus attachment protein which initiates the first interaction of the virus and the infected cells. Antibody capable of binding with the protein will be likely preventing the viral infection (Barnett et al., 2001). In the case of JTat recombinant protein is a minor protein required by the virus to activate the transcription of the viral genome into mRNA which is necessary for the efficient replication of the virus in the infected host (Cota-Gomez et al., 2002; Wu, 2004). Immune response against the protein is expected to be able to slow down or completely stop the viral replication.

Laboratory scale production was initiated to get a small volume of recombinant proteins which used to field trial in Bali cattle at the disease investigation centre, Denpasar, Bali. Those three recombinant proteins were used in the field trial, and the results gave good response in resulting antibody. Scaling up of the recombinant proteins needs to be attempted through industrial link collaboration.

The relevant of ethical features to the JDV recombinant protein research
In view of the fact that only few articles published on the ethical reflections on molecular technology. In biomedical production, it is rather impossible to be produced conventionally in this recent era. For human needs, increasing population in the world would tend to increase the problems in health due to very rapid development of industries for many purposes. The effect of industrial process, increasing transportation, changing the green view of trees into buildings, all creates pollutant. This implies that the people health decrease and the needs of medicines increase drastically. The availability of medicines would not impossible be produced conventionally. Discoveries in molecular biology technology, it contributes significantly in increasing wider products such as in agriculture, medicine, energy, etc. In agriculture base, introduction of advanced technology in food production (cereal, paddy, corn, etc.) is the most important and very concerned in the world. Increasing of animal population is also important as protein source from livestock. This protein source derived from livestock consists of more complete in number of amino acids compared to that protein source from plant. Similarity to human, population of livestock can tend to decrease due to infected by some diseases or less feed.

Jembrana disease is an acute and severe disease, endemic in Bali cattle (Bos javanicus). The disease caused a 20% case fatality rate after a short incubation period in Bali cattle (Moll, 1998). However, in the later publication the JDV is also infecting to other cattle types and buffalo under experiment and become infected under field condition (Soeharsono et al., 1995). Up to present, preventing to Jembrana disease in Bali cattle has been using crude vaccine for its vaccination (Hartaningsih and Wilcox, 1996; Hartaningsih et al., 2001). The crude vaccine is provided from infected tissue of Bali cattle by Jembrana viruses. Availability of this crude vaccine is limited compared to recent population of Bali cattle in the field. Despite limited in production, the crude vaccine is also costly, less effective and demanding in vaccine maintenance.

Entering biotechnology era, it is possible to develop a conventional vaccine such as Jembrana vaccine by recombinant DNA technology to produce recombinant viral protein as Jembrana vaccine. Development of recombinant protein for potential vaccine was initiated by colleagues at Murdoch University for Jembrana disease virus (JDV). Initiation of producing a recombinant Jembrana vaccine is therefore expected to solve some of the problems associated with disadvantages of the current crude vaccine. This recombinant Jembrana vaccine is a safer, effective and lower cost in producing.

Consideration of using advanced technology has based researchers to develop the existing Jembrana vaccine. They (researchers) should decide independently in choosing the manner of doing their research as long as till in the save corridor of principles of biomedical ethics. Principles of biomedical ethics as stated by Beauchamp and Childress (2001) consist of respect to autonomy (independency in decision-making); justice (fair treatment and equity); beneficence (doing the good thing); and non-maleficent (to do no harm).

Researchers should concern to the tendency of increasing people in the world. Based on Malthus Theory (1798), it was featured that world population increasing exponentially: 2, 4, 8, 16 etc. while food production increasing linearly: 1, 2, 3, 4 etc., then human population in 2002 was 6,137 million (4,944 million are in developing countries) with rate increasing of 1.3% per annum. Therefore, it is no point that we have to do something to meet with those world features in order to get secure life and harmony with nature. The decision of doing something is not easy to implement it to the research as reminded by Abu Ali al-Husain ibn Abdallah ibn Sina (Avicenna), 980-1037 (http://www-history.mcs.st-andrews.ac.uk/Mathematicians/Avicenna.html1990). His statement was “In science, humankind is (still) looking for a harmonious relationship with nature, and seeking to know it and conform to it”

In Science, there are many things to be taken and decided with as much as zero risk to meet with goal of creating a life in harmony. To implement this statement, researcher will use advanced technology as a tool based on molecular biology; some will work on genetically engineered organism or genetically modified organism. Those laboratory works will increase the productivity of food or medicines. However, researchers should always control of their work in using the tool and research material. As reminded by first bioethicist Aldo Leopold (1949) that “A thing is right when it tends to preserve the integrity, stability, and beauty of the biotic community. It is wrong when it tends otherwise”.

The integration of utilitarianism principle to the producing of JDV recombinant vaccine
Concomitant with using a tool wisely, development of Jembrana vaccine was aimed to fulfill the vaccine needs in a large number due to the increase of the bali cattle population. To achieve this goal, there is no point that researchers have to choose the right and a wise tool. A recombinant DNA technology and more precisely recombinant protein technology is an approach to answer the needs of Jembrabna vaccine.

The aim of using the advanced technology is to clone a specific gene target into a plasmid vector, then transformed into an E. coli host cell. In the culture, the host cell of E. coli will be multiplied in numbers then recombinant proteins are expressed in side of the cells. Methods of SDS-PAGE and Western Blotting are used for purification and characterization of the recombinant proteins. The yield of recombinant proteins by implementation of recombinant DNA technology is therefore extremely huge number which this product of recombinant proteins is potential as a Jembrana recombinant vaccine. As stated in the previous chapter, those potential recombinant proteins are JSU, JTm and JTat. Those kinds of recombinant proteins are much more qualified vaccine due to obtained from targeted gene of Jembrana viral genome compared to that of crude vaccine. The crude vaccine is not only expensive in processing due to obtaining from killing the bali cattle to get the infected organs but the crude vaccine is also not targeting. Not targeting, it is because we do not know exactly that the killed cattle due to infected by the single disease of the jembrana virus.

Compared to the conventional crude vaccine, the recombinant vaccine has some advantages. The recombinant vaccine can be produced cheaper in mass production, more qualified (because of more targeting) and more practical in handling to the field. This mass production comes along with the increase of Bali cattle population recently which is not only in Bali Island but spread wider almost throughout of Indonesian islands.

Line up with the processing of recombinant protein production, recombinant vaccine could be produced in a large yield through E. coli. This research act is an implementation of biomedical ethics principles: respect to autonomy, justice, beneficence and non-maleficent (Beauchamp and Childress, 2001). In doing biomedical research, researchers have responsibility upon the products therefore they should be carefully and always remember for some signs of biomedical ethics.

Much medicine products have been processed through GMO (genetically modified organism) or GEO (genetically engineered organism) to get mass production otherwise people in the world would safer from illness. Researchers doing those processes both in the laboratory and the trial have take into account for the safety. At least one of two principles in ethical theory has fundament the resulting from the research processes, this is Utilitarianism principle while another is Deontology principle (responsible and entitle of each individual to another individual), (Bentham, 1907; Mill, 1998). There is an integration of producing recombinant protein in a large number to the Utilitarianism principle. The Utilitarianism principle follows the concept of producing the greatest good for greatest numbers. The greatest good in such matter is recombinant protein product which is the qualified product with targeted immunity while the greatest numbers are meant as a large yield producing from manufacturing of E. coli as a host cell.

Respect of researchers in the laboratory and in the field is expected to create that humankind and other individual live in harmony

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