Professional Documents
Culture Documents
11 Pathogen
11 Pathogen
• Quantitation of
virulence to compare
strains 100
Percent of cells
Virus /mutant Apoptotic Non-apoptotic
Wild type 0.1 99.9
E1b+, RID 0.6 99.4
E1b+, RID 0.2 99.8
E1b-, RID 9.9 90.2
E1b-, RID 87.2 12.8
• Dengue fever/dengue
hemorrhagic fever
• Primary infection - acute, self-
limiting
• Secondary infection - non-
protective antibodies bind and
facilitate entry to monocytes
through Fc receptor
• Causes cytokine release that
leads to hemorrhage, shock and
death
• Ebola/HIV similar affect
• A - T cell proliferation
• B - virus replication
• C - immunization and
adoptive transfer of T cells
to nude mice; infection
with WT (open circle:
control; closed circle:
mutant virus; square: wt
virus)
Coronavirus • Mouse hepatitis virus
neurovirulence • Neurotropic strains - acute
meningoencephalitis then
chronic demyelination;
noneurotropic - acute
meningitis
• Acute phase - virus
replicates in neurons and
glial cells; then low levels
of viral RNA persist in
glial cells and chronic
inflammation
• Hypothesis: cytokine
response of brain immune
cells determines disease
outcome
• Analysis of mRNA levels of cytokines
24 h following infection of astrocytes
with a neurotropic (MHV-A59) and a
nonneurotropic (MHV-2) virus
compared with an uninfected control
culture. The blots of mouse cytokine
array assays are shown. The cytokine
key is as follows: A, colony-stimulating
factor granulocyte; B, gamma
interferon; C, IL-1; D, IL-1ß; E, IL-2; F,
IL-3; G, IL-4; H, IL-5; I, IL-6; J, IL-7;
K, IL-9; L, IL-10; M, IL-11; N, IL-12
p35; O, IL-12 p40; P, IL-13; Q, IL-15;
R, IL-16; S, IL-17; T, IL-18; U,
lymphotoxin B; V, TNF-; W, TNF-ß; X,
GAPDH; Y, ß-actin; Z, bacterial plasmid
(pUC18).
HIV associated dementia (HAD)
Functional evaluation of Tat transactivation (A) expression vectors encoding the isogenic C-Tat
proteins. Differences within the dicysteine motif of these vectors are highlighted.. (B) Transactivation
of LTR-driven GFP expression by different Tat vectors in 293 cells. (C) Transactivation of LTR-
driven SEAP expression by different Tat vectors in 293 cells. SEAP in the culture medium was
quantified on day 1 (open bars) and day 3 (filled bars). (D) Rescue of the Tat-defective virus by
isogenic C-Tat proteins. HLM-1 cells were transfected with different C-Tat variant expression vectors.
Culture supernatants were collected on days 1, 3, 5, and 7 following transfection, and p24 levels in the
culture supernatants were determined. Results of experiments using samples from day 3 are
presented; similar results were observed for samples from other days. Abs, absorbance; -VE, parental
vector.
Taxis assay:
membrane with
monocytes on one
side and test
protein on other
Count cells on
filter
Monocyte migration induced by isogenic Tat proteins. f-MLP peptide was used as a
positive control at 10-7 and 10-8 M concentrations. Tat proteins were used at
concentrations of 100 and 20 ng/ml (12 and 2.4 nM, respectively) as indicated. No
grad, wells with 100 ng of CC-Tat protein/ml in both the compartments. Differences in
the numbers of monocytes that migrated with Tat-CC and Tat-CS were statistically
significant
Viruses and multiple sclerosis?