Article Figures & Data
Figures
- FIGURE 1.
Pathologic lesions in tissues of 8-wk-old germfree IL-2−/− mice. The lungs (a), liver (b), pancreas (c), kidney (d), small intestine (e), colon (f), and spleen (g) of IL-2−/− mice were fixed in formalin and embedded in paraffin, and 5-μm sections were counterstained with hematoxylin-eosin before photomicroscopy. Note the extensive lymphocyte hyperplasia in all tissues with the exception of the intestine. Magnification: a through e and g, ×200; f, ×300.
- FIGURE 2.
Accumulation of CD4−CD8+ and B220+TCRlow/int intrahepatic lymphocytes in germfree IL-2−/− mice. Representative FACS profile of CD4, CD8, TCRαβ, and B220 expression by cells recovered from the livers of 5-wk-old germfree (GF) IL-2−/− and IL-2+/+ mice. The total number of cells recovered from the liver of each animal is shown above the dot plots. The boxed region in the lower dot plots identifies the B220+TCRαβlow/int population of intrahepatic lymphocytes. The results shown are representative of those obtained from two pairs of mice.
- FIGURE 3.
Phenotypic analysis of spleen cells from 8-wk-old germfree IL-2−/− mice. A, Representative FACS profile of splenic T cells and histogram profiles of CD69 and CD25 expression by CD4+ T cells. The total number of cells recovered from the spleen of each animal is shown above the dot plots. B, Expression of B220 among spleen cells from wild-type (IL-2+/+) and IL-2-deficient (IL-2−/−) germfree mice. The results shown are representative of those obtained from four pairs of mice.
- FIGURE 4.
Cellular composition of bone marrow from 8-wk-old germfree IL-2−/− mice. Representative FACS profiles of bone marrow mononuclear cells from IL-2−/− mice and wild-type (IL-2+/+) littermates using lineage-specific Abs to identify myeloid (Gr-1), monocytic/macrophage (Mac-1), and B (B220) cells. The values shown for Gr-1 profiles represent the frequency of mature, Gr-1high, granulocytes. Those shown for Mac-1 and B220 represent the relative proportions of immature (Mac-1low; B220low) and mature (Mac-1high; B220high) cells present.
- FIGURE 5.
Flow cytometric analyses of iIEL from 8-wk-old germfree IL-2+/+ and IL-2−/− mice. The iIEL were stained with the following combinations of Abs; anti-Thy 1.2, anti-TCRαβ, and anti-TCRγδ Abs for three-color analysis and anti-TCRγδ or anti-TCRαβ combined with anti-CD4, anti-CD8α, and anti-CD8β Abs for four-color analysis. Viable cells were analyzed by flow cytometry as described in Materials and Methods. The numbers on each dot plot and histogram represent the percentage of total cells present in each quadrant and in the gated areas, respectively.
- FIGURE 6.
Flow cytometric analyses of iIELs from IL-2−/− bone marrow chimeras. The iIELs recovered from chimeras 90 days after reconstitution with either host (IL-2+/+) bone marrow (A) or congenic IL-2−/− bone marrow (B) were stained with anti-TCRαβ and anti-TCRγδ Abs in conjunction with anti-CD45.1 or CD45.2 Abs and analysed by flow cytometry as described in Materials and Methods. The proportions (percentage) of donor-derived (CD45.2+) or host-derived (CD45.1+/CD45.2−) cells present are shown in the upper histograms in A and B. The proportions (percentage) of donor-derived and host-derived TCRαβ+ and TCRγδ+ iIELs shown in the lower histograms in A and B were determined by electronically gating on CD45.2+ or CD45.2− cells. The results shown are representative of those obtained from four animals reconstituted with host bone marrow and five animals reconstituted with IL-2−/− bone marrow.
Tables
Age (weeks) IL-2 Genotype n RBC (106/μl) HB (mg/dl) HT WBC (103/μl) Differential (×109/L) Neut Lymph Mono Eosin 3–5 +/+ 5 7.9 ± 0.9 15.0 ± 0.5 33 ± 1 7.6 ± 1.5 0.6 ± 0.2 6.2 ± 1.1 0.1 ± 0.1 0.08 ± 0.02 +/− 3 7.2 ± 1.2 14.8 ± 0.3 32 ± 2 6.8 ± 2.0 0.7 ± 0.2 6.4 ± 1.3 0.2 ± 0.1 0.09 ± 0.03 −/− 4 7.4 ± 0.8 13.1 ± 1.1 28 ± 5 8.0 ± 2.6 0.3 ± 0.2b 7.0 ± 1.4 0.1 ± 0.1 0.1 ± 0.04 8 +/+ 3 8.6 ± 0.2 14.2 ± 0.7 31 ± 1 11.0 ± 1.9 1.7 ± 0.3 9.7 ± 2.5 0.2 ± 0.2 0.1 ± 0.06 +/− 6 8.9 ± 0.9 15.6 ± 1.0 31 ± 2 10.1 ± 2.1 1.6 ± 0.3 8.6 ± 2.6 0.2 ± 0.1 0.1 ± 0.02 −/− 4 3.8 ± 0.6 10.0 ± 0.5 19 ± 6 9.8 ± 2.5 0.6 ± 0.6c 6.3 ± 2.2b 0.6 ± 0.2 0.2 ± 0.05 12 +/− 4 9.9 ± 1.9 14.9 ± 1.8 33 ± 4 13.1 ± 2.1 1.8 ± 0.3 11.0 ± 3.0 0.2 ± 0.1 0.2 ± 0.1 −/− 7 2.6 ± 2.5 3.7 ± 2.3 12 ± 3 14.0 ± 2.2 0.2 ± 0.2d 5.9 ± 2.0b 0.6 ± 0.2 0.3 ± 0.2 -
a HB, hemaglobin; HT, hematocrit; WBC, white blood cells. Data represent the mean ± SD. Manual 300-count leukocyte differentials were performed on blood smears from sex-matched littermates. Data represent the mean ± SD.
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b p < 0.01 compared with +/+ mice.
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c p < 0.005 compared with +/+ mice.
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d p < 0.0005 compared with +/+ mice.
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b Cells consisted of atypical blast-like cells that were tartrate sensitive, acid phosphatase-positive, and myeloperoxidase-negative.
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Age (weeks) IL-2 Genotype n Splenomegalyb Colitisc Bone Marrowa Spleena Total (×106) % B220+ % CD3+ Myeloid Cells Immatured Maturee Total (×106) % B220+ % CD3+ 4 +/+ 3 − − 51 ± 7 30 ± 5 3 ± 1 31 ± 6 67 ± 8 25 ± 10 43 ± 4 22 ± 5 −/− 4 − − 33 ± 8 33 ± 6 2 ± 2 56 ± 5 40 ± 7 28 ± 7 40 ± 10 25 ± 6 6 +/+ 4 − − 60 ± 10 33 ± 4 8 ± 2 66 ± 4 28 ± 8 35 ± 8 49 ± 5 34 ± 3 −/− 4 +/++ − 24 ± 6 21 ± 9 10 ± 3 84 ± 10 13 ± 5 58 ± 15 35 ± 9 23 ± 6 8 +/+ 3 − − 59 ± 5 29 ± 7 7 ± 1 66 ± 10 31 ± 9 47 ± 17 47 ± 6 37 ± 6 −/− 5 ++ − 19 ± 7 16 ± 4 18 ± 4f 83 ± 12 18 ± 6 73 ± 21 16 ± 4 21 ± 5 -
a Phenotype and composition of bone marrow and spleen cells determined by flow cytometry. Data represent the mean ± SD.
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b Size of IL2−/− spleen relative to that of IL2+/+ mice; −, no change; +, 1 to 2 times larger; ++, >2 times larger.
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c Based upon histologic analysis.
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d Gr-1low and/or Mac-1low cells.
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e Gr-1high cells.
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f 60–75% TCRαβ+CD4+CD69+ and/or CD25+.
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Age (weeks) IL-2 Genotype n RBC (106/μl) HB (mg/dl) HT WBC (103/μl) Differential (×109/L) Neutrophil Lymph Mono Eosin 3–5 +/+ 5 7.9 ± 0.9 15.0 ± 0.5 33 ± 1 7.6 ± 1.5 0.6 ± 0.2 6.2 ± 1.1 0.1 ± 0.1 0.08 ± 0.02 +/− 3 7.2 ± 1.2 14.8 ± 0.3 32 ± 2 6.8 ± 2.0 0.7 ± 0.2 6.4 ± 1.3 0.2 ± 0.1 0.09 ± 0.03 −/− 4 7.4 ± 0.8 13.1 ± 1.1 28 ± 5 8.0 ± 2.6 0.3 ± 0.2ab 7.0 ± 1.4 0.1 ± 0.1 0.1 ± 0.04 8 +/+ 3 8.6 ± 0.2 14.2 ± 0.7 31 ± 1 11.0 ± 1.9 1.7 ± 0.3 9.7 ± 2.5 0.2 ± 0.2 0.1 ± 0.06 +/− 6 8.9 ± 0.9 15.6 ± 1.0 31 ± 2 10.1 ± 2.1 1.6 ± 0.3 8.6 ± 2.6 0.2 ± 0.1 0.1 ± 0.02 −/− 4 3.8 ± 0.6 10.0 ± 0.5 19 ± 6 9.8 ± 2.5 0.6 ± 0.6bc 6.3 ± 2.2b 0.6 ± 0.2 0.2 ± 0.05 12 +/− 4 9.9 1.9 14.9 ± 1.8 33 ± 4 13.1 ± 2.1 1.8 ± 0.3 11.0 ± 3.0 0.2 ± 0.1 0.2 ± 0.1 −/− 7 2.6 ± 2.5 3.7 ± 2.3 12 ± 3 14.0 ± 2.2 0.2 ± 0.2cd 5.9 ± 2.0b 0.6 ± 0.2 0.3 ± 0.2 -
a HB, hemaglobin; HT, hematocrit; WBC, white blood cells. Data represent the mean ± SD. Manual 300-count leukocyte differentials were performed on blood smears from sex-matched littermates. Data represent the mean ± SD.
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bp < 0.01 compared with +/+ mice.
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cp < 0.005 compared with +/+ mice.
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dp < 0.0005 compared with +/+ mice.
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b Cells consisted of atypical blast-like cells that were tartrate sensitive, acid phosphatase-positive and myeloperoxidase-negative.
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Age (weeks) IL-2 Genotype n Splenomegalyb Colitisc Bone Marrow* Spleena Total (×106) % B220+ % CD3+ Myeloid Cells Total (×106) % B2202 % CD3+ Immatured Maturea 4 +/+ 3 − − 51 ± 7 30 ± 5 3 ± 1 31 ± 6 67 ± 8 25 ± 10 43 ± 4 22 ± 5 −/− 4 − − 33 ± 8 33 ± 6 2 ± 2 56 ± 5 40 ± 7 28 ± 7 40 ± 10 25 ± 6 6 +/+ 4 − − 60 ± 10 33 ± 4 8 ± 2 66 ± 4 28 ± 8 35 ± 8 49 ± 5 34 ± 3 −/− 4 +/++ − 24 ± 6 21 ± 9 10 ± 3 84 ± 10 13 ± 5f 58 ± 15 35 ± 9 23 ± 6 8 +/+ 3 − − 59 ± 5 29 ± 7 7 ± 1 66 ± 10 31 ± 9 47 ± 17 47 ± 6 37 ± 6 −/− 5 ++ − 19 ± 7 16 ± 4 18 ± 4a 83 ± 12 18 ± 6 73 ± 21 16 ± 4 21 ± 5 -
a Phenotype and composition of bone marrow and spleen cells determined by flow cytometry. Data represent the mean ± SD.
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b Size of IL2−/− spleen relative to that of IL2+/+ mice; −, no change; +, 1 to 2 times larger; ++, >2 times larger.
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c Based upon histologic analysis.
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d Gr-1low and/or Mac-1low cells.
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e Gr-1high cells.
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f 60–75% TCRαβ+CD4+CD69+ and/or CD25+.
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