-
- Take a good look...read what your tax dollars have patented.
YOU own this.
- Imagine it being used on the American public or any people
on the planet
- as a weapon of mass illness and death. Imagine...
- ------------------------------------------------------------------------
- United States Patent 5,242,820
- Lo September 7, 1993
- ------------------------------------------------------------------------
- Pathogenic mycoplasma
- Abstract
-
- The invention relates to a novel pathogenic mycoplasma
isolated from
- patients with Acquired Immune Deficiency Syndrome (AIDS)
and its use in
- detecting antibodies in sera of AIDS patients, patients
with AIDS-related
- complex (ARC) or patients dying of diseases and symptoms
resembling AIDS
- diseases. The invention further relates to specific DNA
sequences,
- antibodies against the pathogenic mycoplasma, and their
use in detecting
- DNA or antigens of the pathogenic mycoplasma or other
genetically and
- serologically closely related mycoplasmas in infected
tissue of patients
- with AIDS or ARC or patients dying of symptoms resembling
AIDS diseases.
- The invention still further relates to a variety of different
forms of
- vaccine against mycoplasma infection in humans and/or
animals.
- ------------------------------------------------------------------------
- Inventors: Lo; Shyh-Ching (Potomac, MD)
- Assignee: American Registry of Pathology (Washington,
DC)
- Appl. No.: 710361
- Filed: June 6, 1991
-
- Current U.S. Class: 435/252.1; 435/5; 435/872
- Intern'l Class: C12N 005/00; C12N 005/02; C12N 001/00;
C12Q 001/70
- Field of Search: 435/870,5,872,240.2
- ------------------------------------------------------------------------
- References Cited [Referenced By]
-
- Other References
-
-
- Marquart et al (1985) Mycoplasma-Like Structures . .
. Eur J Clin Microbiol
- 4(1):73-74.
- Lo et al (1989) A Novel Virus-like Infectious Agent .
. . Am J Trop Med Hyg
- 40(2):213-226.
- Lo et al (1989) Identification of M Incognitus . . .
Am. J. Trop-Med. Hyg
- 41(5):601-616.
- Lo et al (1989) Association of the Virus-like Agent .
. . Am J Trop Med Hyg
- 41(3):364-376.
- Lo et al (1989) Fatal Infection of Silvered Leaf Monkeys
. . . Am. T Trop
- Med Hyg 40(4):399-409.
- Lo et al (1989) Virus-like Infectious Agent . . . Am
J Trop Med Hyg
- 41(5):586-600.
- Marquart et al (Feb. 1985) Abstract Only Eur J Clin Microbiol
4(1):73-74.
- Hu et al (1990) Gene 93:67-72.
- Primary Examiner: Nucker; Christine M.
- Assistant Examiner: Preston; D. R.
- Attorney, Agent or Firm: Venable, Baetjer, Howard &
Civiletti
- ------------------------------------------------------------------------
- Goverment Interests
- ------------------------------------------------------------------------
-
- The invention described herein was made in the course
of work under a grant
- or award from the United States Department of the Army.
- ------------------------------------------------------------------------
- Parent Case Text
- ------------------------------------------------------------------------
-
- CROSS-REFERENCE TO RELATED APPLICATIONS
- This is a continuation-in-part of U.S. patent application
Ser. No. 265,920,
- filed Nov. 2, 1988, now abandoned, which is a continuation-in-part
of U.S.
- patent application Ser. No. 875,535, filed Jun. 18, 1986,
now abandoned.
- ------------------------------------------------------------------------
- Claims
- ------------------------------------------------------------------------
-
- What is claimed is:
- 1. A biologically pure mycoplasma isolated from tissues
of patients with
- AIDS comprising the mycoplasma produced by the cell line
ATCC No. CRL 9127.
- 2. A biologically pure mycoplasma having the identifying
characteristics of
- M. fermentans incognitus, ATCC 53949.
- ------------------------------------------------------------------------
- Description
- ------------------------------------------------------------------------
-
- BACKGROUND OF THE INVENTION
-
- 1. Field of the Invention
-
- The present invention relates to a novel strain of mycoplasma
isolated from
- a patient with AIDS. The mycoplasma is closely related
to a species of
- human mycoplasma, M. fermentans. Upon characterization
of this mycoplasma,
- it may be classified as a unique strain within the species
M. fermentans
- incognitus.
-
- This novel strain of nycoplasma is referred to hereinafter
as the
- incognitus strain or M. fermentans incognitus.
-
- The invention also relates to use of the mycoplasma M.
fermentans
- incognitus as well as all strains of M. fermentans in
detecting specific
- antibodies in sera of patients with AIDS or an acute
fulminant systemic
- disease and/or animals and its use as a vaccine against
infection by the
- mycoplasma. The invention further relates to incognitus
strain-specific
- antibodies and cross-reactive which later break up into
individual cells
- that are capable of passing through membrane filters
of pore size 0.45
- .mu.m or even 0.22 .mu.m.
-
- A trilaminar cytoplasmic membrane contains sterols, phospholipid
and
- proteins. Therefore, the cells are generally susceptible
to polyene
- antibiotics and to lysis by digitonin.
-
- Replication of the Mycoplasma genome may precede cytoplasmic
division
- resulting in multinucleate filaments before individual
cells are delimited
- by constriction. Budding can also occur.
- Most Mycoplasma species are facultatively anaerobic,
and all known species
- are chemoorganotrophic. The fermentative species of Mycoplasma
utilize
- sugars such as glucose, while non-fermentative species
can utilize
- arginine.
-
- Known mycoplasmas may be grown on complex media, such
as Hayflick medium,
- while fastidious mycoplasmas may be grown on diphasic
SP-4 medium. The
- colonies are usually of the "fried egg" type,
i.e., an opaque, granular
- central region, embedded in the agar, surrounded by non-granular
surface
- growth. The optimal growth temperature of mammalian strains
is
- 36.degree.-37.degree. C.
- Many species of Mycoplasma produce weak or clear haemolysis
which appears
- to be due to the secretion of H.sub.2 O.sub.2. This H.sub.2
O.sub.2
- secretion is believed to be responsible for some aspects
of the
- mycoplasmas' pathogenicity. Known mycoplasmas are commonly
sensitive to
- chloramphenicol and
- tetracyclines.
-
- The Mycoplasma genus currently consists of more than
60 known species which
- are differentiated on the basis of various tests, including
utilization of
- glucose and mannose, arginine hydrolysis, phosphatase
production, the "film
- and spots" reaction and haemadsorption. M. fermentans
antibodies (i.e.
- antibodies to homologous antigenic determinants), including
monoclonal
- antibodies of each, which are useful in detecting incognitus
strain
- antigens in infected tissue of patients or animals. The
invention also
- relates to incognitus strain-specific DNA probes which
are useful in
- detecting incognitus strain genetic materials in infected
tissues of
- patients or animals. Incognitus strain genetic materials
may also be
- detected in infected humans or animals by using specific
incognitus strain
- DNA sequences a homologous M. fermentans DNA sequences
and the polymerase
- chain reaction ("PCR") (U.S. Pat. No. 4,683,202
incorporated herein by
- reference).
-
- The ability to monitor AIDS or other acute fulminant
systemic disease
- status can be of great value. In addition to improving
- prognostication,knowledge of the disease status allows
the attending
- physician to select the most appropriate therapy for
the individual
- patient, e.g. highly aggressive or less aggressive therapy
regimens.
- Because of patient distress caused by more aggressive
therapy regimens, it
- is desirable to distinguish those patients requiring
such therapies. It has
- been found that M. fermentans incognitus is more directly
associated and
- functional deficits of the infected organ systems and
is capable of
- distinguishing such patients.
-
- Mycoplasma is a genus of cell wall-less sterol-requiring,
catalase-negative
- pathogens commonly found in the respiratory and urogenital
tracts of man
- and other animals. The cells of Mycoplasma are typically
non-motile and
- pleomorphic, ranging from spherical, ovoid or pear-shaped
to branched
- filamentous forms.
-
- Filaments are the typical forms in young cultures under
optimal conditions,
- which subsequently transform into chains of coccoid cells
-
- Mycoplasmas are the smallest and simplest free-living
organisms known.
- Mycoplasmas are not obligatory intracellular microorganisms
and are usually
- found extracellularly, but are often found intracellularly
in the infected
- tissues (Mycoplasma, Eds. Wolfgang, J. J., Willette,
H. P., Amos, D. B.,
- Wilfert, C. M., Zinsser Microbiology 19th Ed. 1988, Appleton
and Lange,
- 617-623). The term mycoplasma apparently was first used
by B. Frank in 1889
- (Frank B., Dent. Bot. Ges., 7, 332 (1889) and Krass,
C. J. et al., Int. J.
- Syst. Bacteriol. 23, 62 (1973)). Frank, after careful
microscopic
- observation, began writing about invasion of plants (legume)
by these
- microorganisms and stated: "the changed character
of the protoplasm in the
- cortical cells arising from infection, I will designate
as mycoplasma".
- Later, he had more explicitly defined mycoplasma as a
mixture of small
- fungus-like microorganisms and cell protoplasm (Frank,
B., Landwirt. Jahrb.
- 19, 523 (1890)). The description reflected the difficulty
of
- differentiating this unique microorganism from the infected
host cells
- morphologically.
-
- Even today with electron microscopy, it is still often
difficult to
- differentiate the mycoplasmas from the cellular protoplasmic
processes or
- the subcellular organelles of the infected host, because
ultrastructurally,
- these microorganisms have protoplasm-like internal structures
and are
- bounded by only an outer limited membrane (unit membrane)
without a cell
- wall. Thus, there have been few electron microscopic
studies of mycoplasmas
- identified directly in the infected tissues of animals
or humans.
-
- It has been reported that ultrastructural examination
of infected tissues
- has failed to localize the microbe, even in tissues where
very high titers
- (>10.sup.9 /gm) of microorganisms were recovered in
culture (Elizan, T. S.
- et al., Pro. Soc. Exp. Biol. Med. 139, 52 (1972) and
Schwartz, J. et al.,
- Pro. Soc. Exp. Biol. Med. 139, 56 (1972)). Therefore,
morphologically, the
- microbe might be mimicking certain normal cellular or
subcellular
- structures in the infected host tissues and preventing
direct
- identification.
-
- In addition to the natural difficulty of morphological
differentiation
- between the microorganisms and the protoplasm of infected
cells, the often
- poorly preserved formalin-fixed clinical materials present
further
- limitations to any attempt to directly visualize mycoplasma
organisms in
- the tissues.
-
-
- DESCRIPTION OF THE BACKGROUND ART
-
- Acquired Immune Deficiency Syndrome (AIDS) is a devastating
disease that
- has afflicted over 70,000 people worldwide (AIDS Weekly
Surveillance
- Report--United States, Centers for Disease Control, Aug.
29, 1988). The
- disease is clinically characterized by a set of typical
syndromes which
- manifests itself by the development of opportunistic
infections such as
- pneumocystic carinii pneumonia (PCP), toxoplasmosis,
atypical
- mycobacteriosis and cytomegalovirus (CMV). Further characteristics
of the
- AIDS associated syndromes are the clinical manifestation
of
- neuropsychiatric abnormalities, of AIDS encephalopathy
(Naura, B. A., et
- at., Ann.Neuro 19, 517 (1986)), kidney failure of AIDS
nephropathy, heart
- failure of AIDS cardiomyopathy infections and certain
uncommon malignancies
- such as Kaposi's sarcoma or B-cell lymphoma (Durack,
D. T., N.Eng.J.Med.
- 305, 1465 (1981); Reichert, C. M., et al., Am.J.Path.
112, 357 (1983);
- Ziegler, J. L., et al., N.Eng.J.Med. 311, 565 (1984)).
-
- Through co-cultivation of AIDS patients' peripheral blood
cells with
- mitogen-stimulated normal human lymphocytes or permanent
human T-cell
- lines, a number of laboratories have isolated T-cell-tropic
human
- retroviruses (HTLV-III/LAV), Barre-Sinoussi, F., et al.,
Science 220, 868
- (1983); Gallo, R. C., et al., Science 224, 500 (1984).
Epidemiologically,
- the newly isolated retroviruses have been shown to be
highly associated
- with patients of AIDS and/or AIDS-related complex (ARC).
Schupback, J., et
- al., Science 224, 503 (1984); Sarngadharan, M. G., et
al., Science 224, 506
- (1984). In vitro studies with HTLV-III/LAV have demonstrated
T-cell tropism
- and cytopathic changes. Barre-Sinoussi, F., et al., supra;
Popovic, M., et
- al., Science 224, 497 (1984). HTLV-III/LAV is believed
to be the causative
- agent of AIDS.
-
- However, the establishment of an animal model of AIDS
by HTLV-III-LAV
- injection has not been successful. Gajdusek, D.C., et
al., Lancet I, 1415
- (1984). The chimpanzee is the only primate other than
man found to be
- susceptible to infection by HTLV-III/LAV. However, overt
AIDS manifested by
- the development of opportunistic infections and/or unusual
malignancies has
- not yet been seen, despite evidence for persistent infection
and/or viremia
- in experiments on this species. Gajdusek, D.C., et al.
Lancet I, 55 (1985).
- Thus, the human retroviruses have not fulfilled Koch's
postulates, i.e.,
- producing transmissible AIDS-like diseases in experimental
animals.
- HTLV-III/LAV is not associated with the unusual malignancies
such as B-cell
|