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Technological advances in the acquisition of DNA and protein sequence information and the resulting onrush of data can quickly overwhelm the scientist unprepared for the volume of information that must be evaluated and carefully dissected to discover its significance. Few laborat ...

After more than 20 years of intense research, a safe and effective vaccine against HIV-1/AIDS has not been developed. Passive immunization has been used as a tool to demonstrate the role of neutralizing antibodies in conferring protection against HIV-1 challenge in chimpanzees. Because th ...

As described in Chapter 1, the complement system comprises a battery of at least 20 components capable of immediate response to a foreign organism or cell, resulting in lysis or opsonization and phagocytosis. Uncontroled complement activation would result in damage to host tissues, or in ext ...

Lysis is the final end point of complement attack, indicating that the mechanisms acting to protect the cell have been overwhelmed. There is, of course, enormous biological significance in the deposition of complement fragments on cells in vivo because these serve as opsonins and opsonizat ...

All of the basic functional assays of complement activity utilize erythrocytes as targets (1). For classical pathway assays, the favored target is the antibody-sensitized sheep erythrocyte. For alternative pathway assays, unsensitized rabbit erythrocytes are routinely used. ...

Immunoaffinity protocols offer a rapid, efficient, and simple way of obtaining a protein of interest. Almost all of the complement components, receptors, and regulators have been successfully isolated by immunoaffinity protocols. The use of these methods is limited only by the availab ...

Animal models have been widely used to investigate the role of the complement system in a host of infectious and inflammatory diseases. Over the last 30 yr, a number of naturally occurring genetic deficiencies of complement components have been described in different species of laboratory a ...

Serum sickness resulting from repeated administration of horse antitoxins was hypothesized to be an immune complex disease by von Pirquet and Schick at the turn of this century (1). Arthus developed the first animal model of immune complex disease by inducing cutaneous vasculitis and infl ...

The classical pathway of complement is activated in various diseases. A main inhibitor of this pathway is the serpin C1-inhibitor (C1-Inh), which regulates activation at the level of C1. C1-Inh also inhibits the mannan binding lectin (MBL) associated serine proteinases−1 and −2. A deficiency ...

The aim of this chapter is to describe methods for purification of the individual complement (C) components using classical chromatography methods available in most biochemistry laboratories. None of these methods require the large amounts of specific antibodies needed for the pop ...

Complement regulators and receptors can be identified individually in tissues by probing thin (5 μm) tissue sections fixed on a solid glass support with appropriate antibodies. The bound antibodies are visualized with a proper label that has been attached to the primary antibody itself or to a ...

The complement (C) system consists of a group of 12 soluble plasma proteins that interact with one another in two distinct enzymatic activation cascades (the classical and alternative pathways) and in the nonenzymatic assembly of a cytolytic complex (the membrane attack pathway) (Fig. 1; T ...

Latent infection of B lymphocytes by Epstein-Barr virus (EBV) results in the immortalization of the infected cells (1,2). In vitro, expression of latent membrane protein 1 (LMP1) is essential for the proliferation of EBV-immortalized B cells in that LMP1 simulates an activated CD40 receptor ...

This chapter summarizes the genes and mRNAs that have been mapped on to the B95-8 EBV genome. The complete sequence of this strain of Epstein-Barr Virus (EBV) was established (1,2) and data from many publications has been integrated into the map, which is an update of that published previously (3). The B95 ...

The quantitative, competitive polymerase chain reaction (PCR) assay outlined in this chapter was designed for the detection and quantitation of replicated DNAs in both short-term and long-term assays (1). Quantitative, competitive PCR can be used to study both the contribution of prot ...

The two small virally encoded RNA species, EBER-1 and EBER-2, are abundantly expressed in almost all Epstein-Barr virus (EBV)-infected cell types. Their functions in relation to the physiology of the virus remain enigmatic. In recent years, the main interest in the EBERs has been in connection wi ...

The Epstein-Bar virus (EBV)-encoded complementary strand transcripts (CSTs) are a recently described (1,2) group of related transcripts initiating from a common promoter that exhibit a complex splicing pattern and contain a number of small open reading frames with the potential to enc ...

Upon infection with Epstein-Barr virus (EBV), primary human B-lymphocytes are efficiently immortalized and give rise to lymphoblastoid cell lines in vitro. Four of the 11 viral genes expressed in the immortalized B cells have been found to be essential genetically for the process of immort ...

Herpesviruses have two distinct phases to their life cycle. Characteristically, they persist as a latent infection for the lifetime of the infected host. This usually involves a very small number of infected cells in a particular tissue where the virus is present at very low copy number and there is ...

Fluorescence in situ hybridization (FISH) is the method of choice for visualization of viral nucleotide sequences in the infected cells. FISH methodology has been previously used for localization of Epstein-Barr virus (EBV) DNA sequences within interphase nuclei or on chromosomes ( ...