Beruflich Dokumente
Kultur Dokumente
B, 1-11
Robin A. Weiss
Introduction
Retroviruses infect a wide variety of vertebrates ranging from fish to humans (Weiss
et al., 1985; Levy, 1992-4). Some retroviruses cause diseases (Table I) of agricultural
and economic importance, e.g. leukosis of cattle and chickens. Of course, the greatest
impact has come from AIDS, a disease which was not recognised until 1981. Human
immunodeficiency virus (HIV) is the topic of this symposium and this article is to
introduce retroviruses in general and to explore their interactions with host cells.
Retrovirus replication
Retroviruses acquired their name when the discovery in 1970 of reverse transcriptase
(RNA dependent DNA polymerase) showed that their RNA genomes are copied into
DNA. This RNA to DNA conversion was the first example of a 'backwards' direction
of genetic information in biological systems. It has formed the basis of the first clinically
applied antivirals for retroviruses, because drugs such as azidothymidine, dideoxy-
cytidine and lamivudine (3TC) are selectively incorporated as DNA chain terminators
by viral reverse transcriptase and not by host DNA polymerases. Now we know that
a broader group of genetic elements than retroviruses utilise reverse transcription at
some stage of replication; these include hepadnaviruses (including hepatitis B virus),
cauliflower mosaic virus and retrotransposons of eukaryotes and prokaryotes. Indeed
lamivudine may find a place in the treatment of hepatitis B infection as well as HIV.
A simplified version of the replication cycle of a retrovirus is shown in Figure 1. The
virus particle is diploid, i.e. it carries two genome copies in its core. Reverse transcriptase
is also packaged in the core but does not become enzymatically active until penetration
i
03O5-7453/96/37BO01 + 11 $12.00/0 t . 1996 The British Society for Antimicrobial Chemotherapy
R. A. Weiss
Malignancies
myeloid leukaemia
erythroid leukaemia
lymphoid leukaemia
lymphoma
sarcoma
mammary carcinoma
renal carcinoma
Haematological deficiencies
aplastic anaemia
haemolytic anaemia
autoimmune disease
immunodeficiency
Bone and joint disease
and uncoating of the virion takes place. The primers for reverse transcription are
cellular transfer RNA molecules that hybridize with the viral RNA genome, tRNAlys
for HIV. From these primers nascent DNA chains are formed. Reverse transcriptase
also has an RNaseH activity that removes the parental RNA from the DNA, which
then forms the template for second strand DNA synthesis to form a double-stranded
DNA 'provirus'. This provirus is haploid but has sequences derived from both RNA
Receptor
Retrovirus classification
Retroviruses are broadly classified into three subfamilies, oncoviruses, lentiviruses and
spumaviruses (Table II), based on electron microscopic morphology of virions and
analysis of genome sequences. Oncoviruses are further subclassified into B- C- and
D-type particles morphologically (A-type particles are cytoplasmic cores of B- and
D-type particles). Not all retroviruses classed as oncoviruses are oncogenic. Some are
apparently harmless in their natural hosts; others such as the D type simian retroviruses
related to Mason-Pfizer monkey virus induce an immune deficiency which can be as
severe as those caused by HIV and SIV. Foamy viruses (spumaviruses) frequently occur
in monkeys and great apes, especially in the central nervous system, but the status of
human foamy virus has become uncertain. They are not known to be pathogenic in vivo,
despite the fact that they are more cytopathic in vitro than any other retroviruses,
forming large, vacuolated syncytia with a foamy appearance.
An alternative broad classification of retroviruses that has recently come into
vogue is to divide them according to genome organisation into 'simple' and 'complex'
viruses. Simple retroviruses contain only gag pol and env genes, while complex
retroviruses contain extra non-structural genes having regulatory or auxiliary functions.
The genome organisation of some typical retroviruses is shown in Figure 2. gag,
pol and env encode the core proteins, viral enzymes and envelope glycoproteins
respectively. The regulatory genes include transcriptional activators such as tat of
HIV, tax of human T cell leukaemia virus (HTLV-I) and bel-\ of human foamy virus
(HFV) which upregulate viral transcription. They may also transactivate cellular genes;
for example, tax protein acts indirectly to enhance expression of the p55 chain of
the interleukin-2 receptor which is overexpressed in the T-cell leukaemia induced by
this vims.
HTLV-1
nef
HIV-1
bell
HFV LTR
ML..
kb
6 8 10 12
Figure 2. Proviral genomes of different classes. The simple oncovirus, mouse leukaemia virus (MLV)
possesses only gag, pol and enc genes whereas the complex viruses shown (HTLV-I, H1V-I and HFV) have
accessory genes which aid viral replication. All the genes or their products are potential targets for anti-viral
drugs.
Retrovirus classification and cell interactions S
Retrovinis transmission
Retroviruses are transmitted horizontally, vertically, and genetically as integrated DNA
proviruses in germ cells. HIV can be transmitted horizontally both as a cell-free virus,
or as infected cells, whereas HTLV is only cell-associated. Hence the frequent presence
of both HTLV and HIV in injecting drug users, but only HIV in haemophiliacs (before
screening was introduced).
Horizontal transmission is well adapted to the hosts being infected. Thus some
retroviruses are transmitted through biting and scratching (e.g. leukaemia vims among
cats, SIV among macaques), via water and gill wounds among fish, and via biting flies
among horses in equine infectious anaemia virus (EIAV).
Vertical transmission of infectious virus can occur prenatally, perinatally and
post-natally, as known for HIV. The majority of paediatric infections appear to occur
perinatally but milk transmission also occurs, particularly on primary maternal
Human retroviruses
Table III lists the known human retroviruses. The first 'human' retrovirus to be isolated
in 1971 was human foamy virus (HFV) from a nasopharyngeal carcinoma line.
R. A. Weiss
Virus Disease
Oncovirus
HTLV-I Adult T-cell leukaemia
Tropical spastic paraparesis
HTLV-II Hairy cell leukaemia?
Neurodegeneration?
Lentivirus
HIV-1 Acquired immune deficiency syndrome
HIV-2 Acquired immune deficiency syndrome
Spumavirus
HFV? None
HTLVs may have been present in humans for thousands of generations, whereas the
HIVs appear to be recent introductions during the past few decades.
Humans also harbour endogenous oncoviral genomes as already discussed,
resembling C-type and D-type retroviruses. Recently, an oncovirus has been found in
immunosuppressed bone marrow transplant patients, through screening for recovery of
replication-competent retroviral vectors. There is also some evidence of a virus related
to D-type retroviruses in Sjogren's syndrome and normal salivary gland biopsies. These
candidates for new human retroviruses require much further characterisation. They
serve to remind us that they may be further human retroviruses awaiting discovery.
Retroviral pathogenesis
Retroviruses elicit so many different diseases and symptoms (Table I) that it is not
possible to review them in detail here. However, from the antiviral point of view, it is
tax
chromosomes, the higher the virus load in the target tissue, the higher the chance that
one cell among millions infected will integrate its provirus in such a dangerous locus
that monoclonal malignancy ensues. Studies of leukosis in chickens have shown that
this occurs in B-cells in the bursa of Fabricius during primary infection, so subsequent
antiviral treatment would presumably be of no avail. In mice and cats, however,
multiple recombination of infectiously spreading leukaemia virus with related yet
distinct endogenous envelope genes appear to be required before the typical T-cell
lymphomas are initiated. Hence antiviral therapy should suppress leukaemogenesis. In
fact the best in-vivo mouse model for testing antiretrovirals (it provided the earliest
evidence for azidothymidine) is Friend or Rauscher erythroid leukaemia in which
splenomegaly is rapidly caused by an oncogene-carrying defective virus coupled with
an infectiously spreading helper virus.
Human T-cell leukaemia exerts its oncogenic effect neither by transducing an
oncogene nor by integrating into DNA next to an oncogene. Rather, its tax gene
Quantitative
viral burden
viral dynamics
Qualitative
genome diversity
antigen variation
cellular tropism
virulence
Retrovirus classification and cell interactions
Virus
(and host) Syndrome Cell tropism Receptor
emphasise the great importance of using therapies that will inhibit viral replication.
Indeed, immune restoration therapy may only make things worse by providing more
activated T-lymphocytes for HIV to replicate within (Coffin, 1995), unless accompanied
by antiviral treatment.
Despite these new insights into viral dynamics we still do not know precisely how HIV
causes AIDS (Weiss, 1993). HIV infects T-helper lymphocytes, macrophages and also
dendritic cells (Patterson el al., 1995). Escape from virus neutralization can change
cellular tropism and vice versa (McKnight et al., 1995). Maedi-visna virus ( M W ) , on
the other hand, infects just the antigen-presenting cells. M W disease in sheep resembles
a subset of the AIDS syndrome, namely the CNS disease and wasting disease (Table V).
It seems reasonable, therefore, to attribute the severe immune deficiency seen in HIV
and feline immunodeficiency virus infection to the infection and depletion of T-helper
lymphocytes, and the CNS and wasting syndrome to infection of macrophages,
including the microglia in the brain.
Cell tropism of retroviruses is determined by cell surface receptors allowing binding
and entry, and further by tissue-specific transcriptional regulation acting on the proviral
LTR. Retroviruses use diverse receptors such as CD4 by HIV and SIV, amino-acid and
phosphate transporter molecules by mammalian C type retroviruses and low-density
lipoprotein related molecules by avian leukosis viruses (Wimmer, 1994). HIV also binds
to the glycolipid, galactosylceramide, which may be relevant to CNS disease. The
distinctive tropism of HIV-1 for T-lymphocytes versus macrophages is determined in
part by the V3 loop of gpl20 which is thought to interact with secondary cellular
receptors following the CD4 interaction (Wimmer, 1994; McKnight et al., 1995).
For the same reason as the limited success of antiviral treatment, vaccines protecting
against lentivirus infection will have to show protection against the myriad
(quasi-species) of substrains present in the infected host populations. One may roughly
calculate that HIV varies over the course of a 10-year infection period within a singly
infected person as much as HTLV-I has done over 100,000 years of co-evolution with
its human hosts worldwide. Whereas a vaccine protective against HTLV-I, based on
env antigens appears quite feasible (as it protects against HTLV-I infection in
experimentally infected rabbits and macaques), the development of HIV vaccines will
be much more problematic.
But that is not to say that efficacious HIV vaccination will be impossible. A small
proportion of African prostitutes exposed to HIV have developed cell-mediated
immunity which appears to protect them although they are repeatedly at sexual risk of
infection (Rowland-Jones et al., 1995). These individuals have never seroconverted to
HIV and lack PCR-detectable HIV genomes in the blood, although they may have been
Therapeutic retroviruses
The foregoing discussion has dealt with naturally occurring retroviruses. It is worth
reminding the reader, too, that retroviruses can be turned to therapeutic use as vectors
in gene therapy. The property of retroviruses in causing persistent infection with genome
integration allows genes to be delivered to cells and tissues. Just as retroviruses have
themselves transduced host oncogenes (Figure 3), so can gene therapists insert genes for
replacement therapy in inherited disorders (Lever & Goodfellow, 1995). Thus, the initial
clinical trials of gene therapy for adenosine deaminase deficiency have used retroviral
vectors derived from disabled murine leukaemia virus genomes. For cancer treatment
the gene for herpes simplex thymidine kinase has been delivered to the tumour so that
systemically administered ganciclovir is selectively activated in the tumour tissue.
Thus it is satisfying to virologists that a viral vector delivering a gene activating an
antiviral prodrug may prove useful in treating a non-viral disease. However, there is
a long way to go to improve both the safety and efficacy of retroviral vectors. This
propensity to integrate is both part of their benefit and a cause of anxiety lest they cause
tumours via insertional mutagenesis. Their delivery is still not sufficient to infect the
majority of the target cells. Improvements are underway to use selective receptors and
tissue-specific gene promotors to restrict expression to target cells (Lever & Goodfellow,
1995). In-vivo delivery is limited by titre and by the observation that human
complement rapidly inactivates murine leukaemia virus envelopes. We have recently
found ways to obviate complement resistance that may lead to a new generation of
retroviral vectors (Takeuchi et al., 1994).
Retro virus classification and cell interactions 11
Acknowledgements
The work of the author is supported by the Medical Research Council and the Cancer
Research Campaign.
References
Note: The retrovirus literature is voluminous. Therefore some key text books and review articles
are cited together with selected articles of topical interest.
Cao, Y., Qin, L., Zhang, L., Safrit, J. & Ho, D. D. (1995). Virological and immunological
characterization of long-term survivors of human immunodeficiency virus type 1 infection.
New England Journal of Medicine 332, 201-8.
Coffin, J. M. (1995). HIV population dynamics in vivo: implications for genetic variation,
pathogenesis, and therapy. Science 267, 483-9.
Ho, D. D., Neumann, A. U., Perelson, A. S., Chen, W., Leonard, J. M. & Markowitz, M. (1995).