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Table 1 A - Three-week but not four-week old BMMC are capable of antigen presentation and B - Loss of APC function between three and four week old BMMC

From: The antigen presentation function of bone marrow-derived mast cells is spatiotemporally restricted to a subset expressing high levels of cell surface FcεRI and MHC II

A

DBA/2 (H-2d)

Culture conditions

Antigenic stimulation (SI)

3Do-54.8

Background (CPM)

4 week old

BMMC, GM-CSF primed

OVA

OVAp

IgE-TNP-OVA

None

 

WEHI-3

0.74

1.1

.95

1,504

 

D11 sup

.86

.76

.58

2,045

 

Rec IL-3

.75

.80

.63

1,786

 

SCF

1.3

.67

.75

1,745

 

WEHI-3+SCF

.95

.87

1.0

3,164

 

WEHI-3+IL-4

.79

.83

.87

2,568

3 week old

BMMC, GM-CSF primed

    
 

WEHI-3

7.8*

8.5*

14.4*

2,359

 

SCF

10.5*

9.6*

15.2*

2,790

 

WEHI-3+IL-4

8.5*

7.4*

22.3*

3,876

B10.BR (H-2 k )

 

Antigenic stimulation (SI)

AD10

None

4 week old

BMMC GM-CSF primed

PCC

PCCP

IgE-TNP-PCC

 
 

WEHI-3 sup

1.4

1.3

0.8

2,140

 

+IL-4

1.2

1.3

.9

1,398

3 week old

BMMC, GM-CSF primed

    
 

WEHI-3

14.4*

9.8*

18.2*

2,575

 

+IL-4

15.2*

12.1*

17.4*

2,132

B

DBA/2 (H-2 d )

Culture conditions

Antigenic stimulation (SI)

3Do-54.8

Background (CPM)

  

Days old of BMMC

GM-CSF primed, in WEHI-3 media

IgE-TNP-OVA

None

  

Day 22

ibid.

10.8*

1,799

  

Day 23

ibid.

12.1*

1,325

  

Day 24

ibid.

14.5*

2,569

  

Day 25

ibid.

12.6*

1,634

  

Day 26

ibid.

3.3*

2,788

  

Day 27

ibid.

0.86

2,961

  
  1. 1A: BMMC (H-2d, or H-2k) were prepared by culturing bone marrow cells in the presence of different sources of IL-3 (20% supernate) or SCF (1 μg/ml) for three or four weeks with weekly decanting of adherent cells. BMMC were primed with GM-CSF (100 U/ml) for 48 hr and some cultures in addition treated with IL-4 at 100 U/ml. BMMC (H-2k) and PCCP-specific AD10 T-cell line were stimulated with pigeon cytochrome c (PCC), PCC peptide (PCCP) or IgE-TNP-PCC complexes at 10 μg/ml for 48 hr and IL-2 production was determined. 1B: For daily assessment of APC function from day 22 to day 27 of the fourth week, nonadherent BMMC were directly obtained from the WEHI-3 supplemented culture wares in the fourth week culture (Day 22 to Day 27) without culture media or plate change. Background CPM was determined in cocultures of differently-treated BMMC plus T cells without antigen stimulation. Stimulation index (SI) was computed as quotients of total CPM of 3H-thymidine uptake of the antigen-stimulated cultures over nonantigen-stimulated controls. Triplicates were set for each group of stimulation and the non-stimulated control, and the means and SE were computed and two-tailed t-test of the antigen-stimulated group compared to background CPM was performed; SE of all observations were calculated within 15% of the computed means. To simplify data, SI with p values < 0.01 was denoted as a * marks in cocultures with competent BMMC stimulation. SI of the group without the mark indicated a failure or loss of antigen presentation by extended cultured mast cells. L-2 secreted in the coculture supernatants was determined by stimulating proliferation and 3H-thymidine uptake of IL-2 dependent NK3 cell line.
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BMC Immunology

ISSN: 1471-2172

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