Immunology

Introduction

Hepatitis B virus is considered to be a major pathogen associated with the chronic and acute hepatitis as well as having an asymptomatic carrier state. It is believed to a great extent that pathogenicity is connected to the immune response that is generated against the viral antigens. I addition to this, it is also observed that very little information is present regarding the T cell response to the envelope antigen. A number of previous studies have mentioned it very clearly that the peripheral cells mediated immune response to HBsAg by the use of crude plasma derive antigen. This is also very important in the pathogenesis and virus clearance. It has also been observed in a study that anti Pre-S2T cell response was also presented before a month of the onset of clinical symptoms. The core focus of this paper is to identify and discuss the different materials and methods used in the experiment regarding the immune response to Hepatitis B virus along with the discussion of the results and findings of the experiment conducted (Mishra, pp. 196).

Materials and Methods

The following are the materials and the people involved in the methods that are included in the conducted experiment regarding the immune response to the Hepatitis B virus.

Patients

It has been observed that seventy three patients were selected with acute hepatitis along with 51 patients with chronic hepatitis. In addition to this, .30 cases of fulminant hepatic failure and seven people who recovered from acute hepatitis B. These people were selected for this experiment with informed consent.

Bacterial Strains and Plasmids

It has been observed that E. coli was obtained from R, Brent and Strain RB 791 was gained as it is a derivative of E. coli K-12 W3110. This involves an F'lac plasmid including a laclQ required for the overproduction of the lac processor. In addition to this, the plasmid pHBV130 consists of coding regions of HBsAg have also been described.

Cloning Reagents and Techniques

It is important to note that transformations, litigations and digestions of the restriction endonuclease were importantly carried out. In addition to this, restriction enzymes along with T4 DNA ligase were purchased from New England Biolabs. In addition to this, the sequences of DNA were determined with the help of chain termination method (Murray, 6284). On the other hand, the Oligonucleotides were synthesized with the help of solid phase phosphotriester method.

Plasmid Constructions

The expression of powerful tac promoter were include for the expression of fusion proteins. This was done for taking advantage of the controlled expression. In addition to this, A part of the lac UV5 promoter was carried by a 509-bp fragment. On the other hand, the coding sequence involving the first 144 residues was removed with the help of pBR1-11 with the digestion of Hpa II. In addition to this, parts of HBsAg, pre-S1 and pre-S2 have also bee carried by the DnA fragments.

Purification of HBcAg Fusion Proteins

Cultures of E. coli strain RB791 involving a variety of plasmids were grown to an OD6w of ...