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Table II.

MHV-68 Latency in the B Cell Compartment During Latent Infection (3 mo after Infection) Is Mainly Maintained in Memory and Germinal Center Cells


Cells

Reciprocal frequency ofgenome-positive cells (SD)a

Percent of total spleenb

Total number of cellsc

Latently infected cellsd
Total B cellse 27,000 (4,600) 42 2.5 × 107 913 
Naive B cellsf 290,000 (150,000) 36 2.2 × 107 74g 
Memory B cellsh 3,100 (2,400) 0.99 5.9 × 105 189 
Germinal center B cellsi
 
1,100 (469)
 
0.44
 
2.6 × 105
 
233
 

Cells

Reciprocal frequency ofgenome-positive cells (SD)a

Percent of total spleenb

Total number of cellsc

Latently infected cellsd
Total B cellse 27,000 (4,600) 42 2.5 × 107 913 
Naive B cellsf 290,000 (150,000) 36 2.2 × 107 74g 
Memory B cellsh 3,100 (2,400) 0.99 5.9 × 105 189 
Germinal center B cellsi
 
1,100 (469)
 
0.44
 
2.6 × 105
 
233
 
a

Frequencies ± 95% confidence limits were determined by linear regression analysis of LDA-PCR data.

b

Percentage of each subset of total spleen cells was determined by FACS® analysis.

c

Total number of cell subset per spleen based on an estimate of 6 × 107 cells/spleen.

d

Number of latently infected cells based on the frequency of viral genome-positive cells within each cell type and its estimated total number per spleen.

e

B cells were sorted as B220+ with 99.17% purity. The contaminating fraction was 0.75% non-B cells.

f

Naive B cells were sorted as IgD/M+ with 99% purity. The contaminating fraction was 0.8% non-B cells and 0.02% isotype-switched cells.

g

The 0.02% contamination with isotype-switched B cells, due to their high level of infection (1 in 2,100), could account for 1 genome-positive cell in every 5,004 purified resting B cells. Therefore, they would make a contribution of one to two cells to the total number (74) of latently infected naive B cells.

h

Memory B cells were sorted as isotype-switched (IgG1+, IgG2ab+, IgG3+, IgA+) CD38high with 95% purity. The contaminating fraction was 2.23% nonisotype-switched B cells and 2.9% germinal center B cells.

i

Germinal center B cells were sorted as isotype-switched (IgG1+, IgG2ab+, IgG3+, IgA+) CD38low with 96% purity. The contaminating fraction was 2.23% nonisotype-switched B cells and 1.9% memory B cells.

Data shown are the mean of three to four independent experiments, each analyzing pooled spleens from five to seven mice. Standard deviation values are shown between brackets.

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