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T cell receptor complementarity determining region 3 length analysis reveals the absence of a characteristic public T cell repertoire in neonatal tolerance. The response in the "tolerant" mouse within the residual repertoire is quantitatively similar but qualitatively different.

Maverakis E, Beech JT, Wilson SS, Quinn A, Pedersen B, Sercarz EE - J. Exp. Med. (2000)

Bottom Line: Our results confirm that despite intraperitoneal injection of neonates with a high dose of HEL emulsified in incomplete Freund's adjuvant, a strong splenic proliferative response to HEL was observed upon recall.After challenge with HEL-complete Freund's adjuvant as adults, immunoglobulin (Ig)G2a isotype antibody was drastically reduced, and IgG1 was found to be the predominant anti-HEL IgG isotype expressed, indicating a deviation of cytokine response toward T helper type 2. 5-wk-old mice, nasally instilled with tolerogenic doses of HEL p106-116, also showed significant inhibition of this public T cell expansion.These results demonstrate that during neonatal and adult nasal tolerance induction, deletion/anergy removes the public clone, exposing a response of similar specificity but that is characterized by the T helper type 2 phenotype and a splenic residence.

View Article: PubMed Central - PubMed

Affiliation: La Jolla Institute for Allergy and Immunology, San Diego, California 92121, USA.

ABSTRACT
All adult BALB/c mice immunized with hen egg white lysozyme (HEL) or its dominant determinant, peptide (p)106-116, mount a T cell response using a "public" Vbeta8.2Jbeta1.5 T cell clone. Neonatal exposure to tolerance-inducing doses of antigen can drastically diminish responsiveness in the draining lymph nodes but not in the spleens of animals challenged as adults with the cognate antigen. To determine the role of T cell deletion or anergy within the mechanisms of observed neonatal "tolerance," we treated neonatal BALB/c mice with HEL and directly followed the characteristic public clone using complementarity determining region 3 length T cell repertoire analysis. Our results confirm that despite intraperitoneal injection of neonates with a high dose of HEL emulsified in incomplete Freund's adjuvant, a strong splenic proliferative response to HEL was observed upon recall. However, the adult splenic T cell response of these neonatally treated mice lacked the usual Vbeta8.2Jbeta1.5 public clone characteristic of HEL-primed BALB/c mice. After challenge with HEL-complete Freund's adjuvant as adults, immunoglobulin (Ig)G2a isotype antibody was drastically reduced, and IgG1 was found to be the predominant anti-HEL IgG isotype expressed, indicating a deviation of cytokine response toward T helper type 2. 5-wk-old mice, nasally instilled with tolerogenic doses of HEL p106-116, also showed significant inhibition of this public T cell expansion. These results demonstrate that during neonatal and adult nasal tolerance induction, deletion/anergy removes the public clone, exposing a response of similar specificity but that is characterized by the T helper type 2 phenotype and a splenic residence.

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Strong splenic proliferative responses persist after neonatal administration of HEL. BALB/c mice received intraperitoneal injections of 50 μl containing 50 μg of HEL emulsified in IFA within the first 18 h of life and again at 72 h. Spleens were harvested at 8 wk of age, and the splenic recall responses to HEL (□) or its dominant determinant, p106–116 (⋄), were measured using a [3H]thymidine incorporation assay. Results are expressed as stimulation indices (S.I.; CPM in sample/CPM in control). In vitro recall responses were significantly elevated (S.I. > 3) for both HEL and p106–116. These data represent the average of three groups of two to three mice.
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Figure 1: Strong splenic proliferative responses persist after neonatal administration of HEL. BALB/c mice received intraperitoneal injections of 50 μl containing 50 μg of HEL emulsified in IFA within the first 18 h of life and again at 72 h. Spleens were harvested at 8 wk of age, and the splenic recall responses to HEL (□) or its dominant determinant, p106–116 (⋄), were measured using a [3H]thymidine incorporation assay. Results are expressed as stimulation indices (S.I.; CPM in sample/CPM in control). In vitro recall responses were significantly elevated (S.I. > 3) for both HEL and p106–116. These data represent the average of three groups of two to three mice.

Mentions: As all HEL-primed BALB/c mice utilize an identical “public” clone that could be readily followed, we measured its expansion in tolerized and untolerized animals to determine whether the mechanism of neonatal tolerance involves T cell deletion or anergy. BALB/c mice were treated neonatally with HEL in IFA (as described in Materials and Methods). Upon reaching adulthood, spleens from neonatally treated animals that had not been challenged in vivo were examined for T cell response after in vitro stimulation with HEL. Fig. 1 shows that adult BALB/c mice treated neonatally with HEL continue to mount a strong in vitro splenic proliferative response to whole HEL as well as to its dominant determinant, p106–116. Clearly, not all HEL-specific T cells are deleted in this form of tolerance induction.


T cell receptor complementarity determining region 3 length analysis reveals the absence of a characteristic public T cell repertoire in neonatal tolerance. The response in the "tolerant" mouse within the residual repertoire is quantitatively similar but qualitatively different.

Maverakis E, Beech JT, Wilson SS, Quinn A, Pedersen B, Sercarz EE - J. Exp. Med. (2000)

Strong splenic proliferative responses persist after neonatal administration of HEL. BALB/c mice received intraperitoneal injections of 50 μl containing 50 μg of HEL emulsified in IFA within the first 18 h of life and again at 72 h. Spleens were harvested at 8 wk of age, and the splenic recall responses to HEL (□) or its dominant determinant, p106–116 (⋄), were measured using a [3H]thymidine incorporation assay. Results are expressed as stimulation indices (S.I.; CPM in sample/CPM in control). In vitro recall responses were significantly elevated (S.I. > 3) for both HEL and p106–116. These data represent the average of three groups of two to three mice.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2195845&req=5

Figure 1: Strong splenic proliferative responses persist after neonatal administration of HEL. BALB/c mice received intraperitoneal injections of 50 μl containing 50 μg of HEL emulsified in IFA within the first 18 h of life and again at 72 h. Spleens were harvested at 8 wk of age, and the splenic recall responses to HEL (□) or its dominant determinant, p106–116 (⋄), were measured using a [3H]thymidine incorporation assay. Results are expressed as stimulation indices (S.I.; CPM in sample/CPM in control). In vitro recall responses were significantly elevated (S.I. > 3) for both HEL and p106–116. These data represent the average of three groups of two to three mice.
Mentions: As all HEL-primed BALB/c mice utilize an identical “public” clone that could be readily followed, we measured its expansion in tolerized and untolerized animals to determine whether the mechanism of neonatal tolerance involves T cell deletion or anergy. BALB/c mice were treated neonatally with HEL in IFA (as described in Materials and Methods). Upon reaching adulthood, spleens from neonatally treated animals that had not been challenged in vivo were examined for T cell response after in vitro stimulation with HEL. Fig. 1 shows that adult BALB/c mice treated neonatally with HEL continue to mount a strong in vitro splenic proliferative response to whole HEL as well as to its dominant determinant, p106–116. Clearly, not all HEL-specific T cells are deleted in this form of tolerance induction.

Bottom Line: Our results confirm that despite intraperitoneal injection of neonates with a high dose of HEL emulsified in incomplete Freund's adjuvant, a strong splenic proliferative response to HEL was observed upon recall.After challenge with HEL-complete Freund's adjuvant as adults, immunoglobulin (Ig)G2a isotype antibody was drastically reduced, and IgG1 was found to be the predominant anti-HEL IgG isotype expressed, indicating a deviation of cytokine response toward T helper type 2. 5-wk-old mice, nasally instilled with tolerogenic doses of HEL p106-116, also showed significant inhibition of this public T cell expansion.These results demonstrate that during neonatal and adult nasal tolerance induction, deletion/anergy removes the public clone, exposing a response of similar specificity but that is characterized by the T helper type 2 phenotype and a splenic residence.

View Article: PubMed Central - PubMed

Affiliation: La Jolla Institute for Allergy and Immunology, San Diego, California 92121, USA.

ABSTRACT
All adult BALB/c mice immunized with hen egg white lysozyme (HEL) or its dominant determinant, peptide (p)106-116, mount a T cell response using a "public" Vbeta8.2Jbeta1.5 T cell clone. Neonatal exposure to tolerance-inducing doses of antigen can drastically diminish responsiveness in the draining lymph nodes but not in the spleens of animals challenged as adults with the cognate antigen. To determine the role of T cell deletion or anergy within the mechanisms of observed neonatal "tolerance," we treated neonatal BALB/c mice with HEL and directly followed the characteristic public clone using complementarity determining region 3 length T cell repertoire analysis. Our results confirm that despite intraperitoneal injection of neonates with a high dose of HEL emulsified in incomplete Freund's adjuvant, a strong splenic proliferative response to HEL was observed upon recall. However, the adult splenic T cell response of these neonatally treated mice lacked the usual Vbeta8.2Jbeta1.5 public clone characteristic of HEL-primed BALB/c mice. After challenge with HEL-complete Freund's adjuvant as adults, immunoglobulin (Ig)G2a isotype antibody was drastically reduced, and IgG1 was found to be the predominant anti-HEL IgG isotype expressed, indicating a deviation of cytokine response toward T helper type 2. 5-wk-old mice, nasally instilled with tolerogenic doses of HEL p106-116, also showed significant inhibition of this public T cell expansion. These results demonstrate that during neonatal and adult nasal tolerance induction, deletion/anergy removes the public clone, exposing a response of similar specificity but that is characterized by the T helper type 2 phenotype and a splenic residence.

Show MeSH
Related in: MedlinePlus