We are sorry, but NCBI web applications do not support your browser and may not function properly. More information

Results: 5

1.
Figure 1

Figure 1. Pancreas histology of protein-immunized and DNA-immunized mice. From: Homing of GAD65 specific autoimmunity and development of insulitis requires expression of both DQ8 and human GAD65 in transgenic mice.

Frozen sections of pancreas were stained with hematoxylin and eosin (A-D) or immune-stained for CD4 (E) or CD8 (F). (A), DNA-immunized non-transgenic mouse pancreas, (B), protein-immunized DQ8-RIP7-hGAD65, double-transgenic mouse pancreas. (C-D), DNA-immunized DQ8-RIP7-GAD65 double-transgenic mouse pancreas. Note the more aggressive characteristic of the lymphocytic infiltration in C (some islet tissue preserved), and D (complete islet destruction) of the GAD65 DNA-immunized double-transgenic as compared with the peri-insular infiltration of protein-immunized double-transgenic mouse (B). Confluent nuclei in E and F (red) correspond to islet area (arrow delineated). Islets are indicated with arrowheads.

Raya B. Elagin, et al. J Autoimmun. ;33(1):50-57.
2.
Figure 2

Figure 2. Summary of islet infiltrate scores following GAD65 DNA or protein immunization protocols in double-transgenic (DT), DQ8 and RIP7-GAD65, single- transgenic, ST), or non-transgenic control mice. From: Homing of GAD65 specific autoimmunity and development of insulitis requires expression of both DQ8 and human GAD65 in transgenic mice.

Insulitis scores were rated as, Grade 0, no insulitis (white bars); grade 1, peri-insulitis (dotted bars); grade 2, insulitis in <50% of the islet (stripped bars); grade 3, insulitis in >50% of the islet (black bars) [39]. The percentage of islets in each scoring category is shown. All 10 DT, DNA-immunized animals developed Grade 2 and 3 insulitis. Only two of ten protein-immunized DT animals developed Grade 2 insulitis and one these mice had both grade 2 and 3 insulitis. All other groups had sporadic or no Grade 2 or 3 insulitis. * Indicates statistically significant differences (p ≤ 0.005) within the same grade, as compared to controls and all other groups.

Raya B. Elagin, et al. J Autoimmun. ;33(1):50-57.
3.
Figure 5

Figure 5. Progression of insulitis in DNA-immunized double-transgenic mice. From: Homing of GAD65 specific autoimmunity and development of insulitis requires expression of both DQ8 and human GAD65 in transgenic mice.

Frozen pancreatic sections, obtained at different time points following DNA immunization, were stained with hematoxilin and eosin (A-E and I), anti-BrdU (F), anti-CD4 (G), or anti-CD8 (H). White arrowheads indicate islets. A, eleven days post-immunization (infiltration present). B, week 5 post-immunization pancreas appeared massively infiltrated. C, week 9 post-immunization (more localized infiltration noted). D, week 13 post-immunization specimen shows a severely infiltrated islet. E: panoramic view of D showing the concomitant presence of normal islet (top left corner). F, post-BrdU treatment of an animal euthanized at week 17 (proliferating cells are exclusively detected in the islet). G and H, pancreas 25 weeks post-immunization (G=anti-CD4, H=anti-CD8). I, pancreas section of a recipient of adoptively transferred splenocytes 5 weeks post transfer.

Raya B. Elagin, et al. J Autoimmun. ;33(1):50-57.
4.
Figure 4

Figure 4. Cellular immunity in immunized mice. From: Homing of GAD65 specific autoimmunity and development of insulitis requires expression of both DQ8 and human GAD65 in transgenic mice.

T-cell proliferation assays were performed using peripheral blood mononuclear cells (PBMCs, left panel) and splenocytes (right panel). DNA-immunized animals’ results shown. Radioactivity from radiolabeled thymidine incorporation to proliferating T cells is expressed in CPM × 103 (Y axis, left margin). DT indicates double transgenics (DQ8-RIP7-hGAD65) while ST indicates single transgenics (GAD65 ST shown). None = no antigen added (negative control), Anti-CD3 was used to stimulate proliferation (positive control). Mean and standard deviation of triplicate results from 5 animals in each group (double or single transgenics) is shown. All double-transgenic animals tested had significant central (splenocytes) antigen specific proliferative responses to hGAD65 (top right) when compared to single-transgenic mice (bottom right).

Raya B. Elagin, et al. J Autoimmun. ;33(1):50-57.
5.
Figure 3

Figure 3. Humoral immunity in immunized mice. From: Homing of GAD65 specific autoimmunity and development of insulitis requires expression of both DQ8 and human GAD65 in transgenic mice.

GAD65 antibodies in DNA or protein immunized mice. Animals were named by a number and the first initial of their gender (f or m). A world standard control for measurement of GAD65 antibodies (GAD6 mMAb) was used as a positive control in all immunoprecipitation experiments. The results were transformed to reactivity units (RU) by defining the maximum amount of GAD65 radioactivity precipitated by GAD6 mMAb as 100%. The experiments were carried out in duplicate and repeated at least three times. The antibody levels of hGAD65 protein-immunized animals (right panel) were consistently higher than those of the hGAD65 DNA-immunized group (left panel). Not all animals developed detectable antibodies in either group (5 of 10 in DNA-immunized vs. 7 of 10 in protein-immunized groups).

Raya B. Elagin, et al. J Autoimmun. ;33(1):50-57.

Supplemental Content

Recent activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...
Write to the Help Desk