A Study to Determine the Immunogenicity and Oral Tolerance to Keyhole Limpet Hemocyanin (KLH)

• ClinicalTrials.gov Identifier: NCT01489956
• Recruitment Status: Terminated
• Results First Posted: April 12, 2016
• Last Update Posted: April 12, 2016
• Sponsor: National Institute of Allergy and Infectious Diseases (NIAID)
• Collaborator: Immune Tolerance Network (ITN)
• Information provided by (Responsible Party): National Institute of Allergy and Infectious Diseases (NIAID)

Tracking Information

• First Submitted Date: December 1, 2011
• First Posted Date: December 12, 2011
• Results First Submitted Date: December 24, 2015
• Results First Posted Date: April 12, 2016
• Last Update Posted Date: April 12, 2016
• Study Start Date: December 2011
• Actual Primary Completion Date: April 2013 (Final data collection date for primary outcome measure)

Current Primary Outcome Measures (submitted: March 11, 2016)

• Participants With a Positive Immune Response to T Cell Stimulation Index (SI) as Measured by 3H-thymidine Incorporation After in Vitro KLH Stimulation of PBMC (Part A) [ Time Frame: Day 16 ]
  T cell stimulation index (SI) as measured by 3H-thymidine incorporation after in vitro keyhole limpet hemocyanin (KLH) stimulation of peripheral blood mononuclear cells (PBMC). An SI ≥3 on day 16 will indicate the presence of immune response. The SI is the ratio of 3H-thymidine incorporation by T cells in the presence of KLH stimulation to 3H-thymidine incorporation by T cells in the absence of stimulation. Higher values correspond with lower tolerance to KLH.
• Participants Demonstrating Tolerance to KLH Using T Cell Stimulation Index (SI) as Measured by 3H-thymidine Incorporation After in Vitro KLH Stimulation of PBMC (Part B) [ Time Frame: Day 32 ]
  T cell stimulation index (SI) as measured by 3H-thymidine incorporation after in vitro keyhole limpet hemocyanin (KLH) stimulation of peripheral blood mononuclear cells (PBMC). An SI <3 on Day 32 indicated tolerance to KLH. The SI is the ratio of 3H-thymidine incorporation by T cells in the presence of KLH stimulation to 3H-thymidine incorporation by T cells in the absence of stimulation. Higher values correspond with lower tolerance to KLH.
• T Cell Stimulation Index (SI) as Measured by 3H-thymidine Incorporation After in Vitro KLH Stimulation of PBMC (Part A) [ Time Frame: Day 9 ]
  No data available for analyses
• T Cell Stimulation Index (SI) as Measured by 3H-thymidine Incorporation After in Vitro KLH Stimulation of PBMC (Part A) [ Time Frame: Day 16 ]
  No data available for analyses.

Original Primary Outcome Measures (submitted: December 9, 2011)

• T cell stimulation index (SI) as measured by 3H-thymidine incorporation after in vitro KLH stimulation of PBMC (Part A) [ Time Frame: Day 0 ]
  An SI>=3 after the booster immunization, day 16, will indicate the presence of immune response
• T cell stimulation index (SI) as measured by 3H-thymidine incorporation after in vitro KLH stimulation of PBMC (Part B) [ Time Frame: Day 42 ]
  An SI <3 after the booster immunization will indicated tolerance.
• T Cell Stimulation Index (SI) as Measured by 3H-thymidine Incorporation After in Vitro KLH Stimulation of PBMC (Part A) [ Time Frame: Day 9 ]
• T Cell Stimulation Index (SI) as Measured by 3H-thymidine Incorporation After in Vitro KLH Stimulation of PBMC (Part A) [ Time Frame: Day 16 ]

Current Secondary Outcome Measures (submitted: March 11, 2016)

• Cytokine Secretion Profile of T Cells Stimulated by KLH (Part A) [ Time Frame: Days 0, 9, 16 ]
  No data available for analyses.
• T Cell Stimulation Index Measured by Carboxyfluorescein Diacetate Succinimidyl Ester (CFSE) Staining After KLH Stimulation (Part A) [ Time Frame: Days 0, 9, 16 ]
  No data available for analyses.
• Suppression (or Non-activation) of Cytokine Secretion Profile of T Cells Stimulated by KLH Following Oral Feeding (Part B) [ Time Frame: Day 42 ]
  No data available for analyses.
• Suppression (or Non-activation) of T Cell Stimulation Index Measured by CFSE Staining After KLH Stimulation (Part B) [ Time Frame: Day 42 ]
  No data available for analyses.
• Other Mechanistic Assessments on Archived Serum Samples Like Anti-KLH Antibodies and Secreted Cytokines (Part B) [ Time Frame: 6 months ]
  No data available for analyses.
• Compare the Level of KLH-specific Antibodies in the Serum (Samples From Various Time Points) Between Parts A and B [ Time Frame: 6 months ]
  No data available for analyses.

Original Secondary Outcome Measures (submitted: December 9, 2011)

• Cytokine Secretion Profile of T Cells Stimulated by KLH (Part A) [ Time Frame: Days 0, 9, 16 ]
• T cell stimulation index measured by carboxyfluorescein diacetate succinimidyl ester (CFSE) staining after KLH stimulation. (Part A) [ Time Frame: Days 0, 9, 16 ]
• Suppression (or non-activation) of cytokine secretion profile of T cells stimulated by KLH following oral feeding.(Part B) [ Time Frame: Day 42 ]
• Suppression (or non-activation) of T cell stimulation index measured by CFSE staining after KLH stimulation.(Part B) [ Time Frame: Day 42 ]
• Other mechanistic assessments on archived serum samples like anti-KLH antibodies and secreted cytokines.(Part B) [ Time Frame: 6 months ]
• Compare the level of KLH-specific antibodies in the serum (samples from various time points) between Parts A and B. [ Time Frame: 6 months ]

• Current Other Pre-specified Outcome Measures: Not Provided
• Original Other Pre-specified Outcome Measures: Not Provided

Descriptive Information

• Brief Title: A Study to Determine the Immunogenicity and Oral Tolerance to Keyhole Limpet Hemocyanin (KLH)
• Official Title: Pilot Study to Determine the Immunogenicity of Immucothel® and Oral Tolerance Induction With Biosyn Native KLH in Healthy Subjects (ITN047AI)
• Brief Summary: The purpose of this study is to test the oral tolerance of Keyhole Limpet Hemocyanin (KLH) and to determine if Immucothel by itself is strong enough to trigger the immune response. If not, Immucothel will be tested in combination with an adjuvant to determine if an adequate immune response can be seen.

Detailed Description

One type of normal immune response is called "oral tolerance." This is when the immune system (the body's natural defense system against illness) turns off (e.g. does not respond) to foods or to other proteins that are eaten. Oral tolerance test is done by feeding people a protein and then vaccinating them with the same protein. Oral tolerance occurs if the vaccination does not cause an immune response.

In this study, oral tolerance of Keyhole Limpet Hemocyanin (KLH) will be tested. KLH is a large protein extracted from a mollusk (a sea animal). The 'native KLH' (which is a large version of this protein) formulation will be used for oral feeding. Immucothel (a smaller version of the KLH protein) will be used for vaccination (injection). Immucothel is an investigational vaccine currently used to treat bladder cancers outside of the US.

Since these particular KLH products have never been used in oral tolerance studies, the investigators want to make sure in this pilot study that they will work as expected in healthy participants before studying these two products in patients with auto-immune disorders.

This study will also determine if Immucothel by itself is strong enough to trigger the immune response. If not, Immucothel will be tested in combination with an adjuvant (a substance that can increase the immune response to a protein like KLH) to determine if an adequate immune response can be seen.

This study consists of two parts. Participants will participate for either 39 days (Part A) or 65 days (Part B). Regardless of the group assignment, a safety follow-up phone call will occur 6 months after the last immunization (189 day for Part A or 215 day for part B) to assess the late onset of adverse events.

Part A of the study will test the experimental vaccine Immucothel by itself or in combination with an adjuvant. Immucothel is a purified protein from a mollusk. Immucothel can be given as a sub-q injection (under the skin) alone or with an adjuvant (a small amount of mineral oil) to help to enhance the immune response. There maybe two groups in this part:

1. Ten evaluable (as defined by protocol) participants will be given Immucothel alone by injection on two occasions, If Immucothel alone creates an immune response in most of the participants then Part A will be completed.
2. If Immucothel alone does not create an immune response in most of the participants in Part A, then 10 new evaluable (as defined by protocol) participants will be asked to volunteer to test Immucothel in combination with the adjuvant Montanide (mineral oil). This will be given by injection on two occasions If there is an immune response in most of the participants to this combination of Immucothel and Montanide then Part A will be done.

Part B of the study will test the successful Immucothel regimen from Part A with oral KLH. Ten new evaluable (as defined by protocol) participants will be given the experimental oral KLH.

• Study Type: Interventional
• Study Phase: Early Phase 1
• Study Design: Allocation: Non-Randomized; Intervention Model: Single Group Assignment; Masking: None (Open Label); Primary Purpose: Treatment
• Condition: Autoimmune Disorders

Intervention

• Biological: Immucothel Alone (Part A)
  100 ug at day 0 (priming dose) and day 9(booster dose)
  Other Names:

  • keyhole limpet hemocyanin (KLH)
  • Low Molecular Weight (LMW) KLH
• Biological: Immucothel+Montanide (Part A)
  100 ug SQ Immucothel plus Montanide at day 0 (priming dose) and day 9 (booster dose)
  Other Names:

  • Immucothel® + Montanide ISA 51
  • LMW KLH +oil based adjuvant
• Biological: Immucothel Alone or Immucothel+Montanide (Part B)
  50 mg of native KLH on days 0-4 and 10-14 (total of 500 mg). Immunization on days 26 and 35.
  Other Names:

  • Immucothel® or Immucothel® + Montanide ISA 51
  • LMW KLH or LMW KLH +oil based adjuvant

Study Arms

• Experimental: Immucothel alone (Part A)
  100 µg Immucothel subcutaneously (SQ) on Day 0 and Day 9.
  Intervention: Biological: Immucothel Alone (Part A)
• Experimental: Immucothel+Montanide (Part A)
  If an immune response was not observed in at least nine out of the first 10 participants after receiving Immucothel alone, 10 additional healthy subjects would be recruited and immunized with Immucothel (SQ) plus Montanide (SQ) on Day 0 and Day 9.
  Intervention: Biological: Immucothel+Montanide (Part A)
• Experimental: Immucothel alone or Immucothel+Montanide (Part B)

  Dependent on the results for Part A.

  Briefly: Ten new, healthy participants were to be fed 50 mg of native keyhole limpet hemocyanin (KLH), a protein extracted from a mollusk (a sea animal), on Days 0 through 4 and Days 10 through 14, for a total dose of 500 mg. The participants were then immunized using the strategy that produced an immune response in at least nine out of 10 participants in Part A (Immucothel alone or Immucothel plus Montanide) on Days 26 and 35.

  Intervention: Biological: Immucothel Alone or Immucothel+Montanide (Part B)

• Publications *: Not Provided

Recruitment Information

• Recruitment Status: Terminated
• Actual Enrollment (submitted: December 20, 2013): 19
• Original Estimated Enrollment (submitted: December 9, 2011): 30
• Actual Study Completion Date: April 2013
• Actual Primary Completion Date: April 2013 (Final data collection date for primary outcome measure)

Eligibility Criteria

Inclusion Criteria:

• Healthy
• Ability to give informed consent and comply with study procedures.
• Participant able/willing to hold off receiving prophylactic immunizations (like influenza or pneumococcal vaccines) during the study period.

Exclusion Criteria:

• Use of corticosteroids within 2 weeks prior to screening visit.
• First degree relative (parent, sibling or child) with history of autoimmune disease.
• Presence of chronic medical illness including but not limited to chronic kidney-, liver-, cardio-vascular diseases, immunodeficiencies, anemia, B12 deficiency, malignancies, or chronic active infections.
• History of acute gastrointestinal illness within 2 weeks prior to oral KLH administration.
• For women of child bearing age, participant unwilling to defer pregnancy, has a positive urine pregnancy test or is currently pregnant or lactating.
• Use of an investigational drug within 3 months of the screening visit.
• History of acute febrile illness within 1 week of screening visit.
• History of allergy to shellfish, previous exposure to KLH/product containing KLH or known-sensitivity to KLH / components of KLH preparation.
• Participants receiving any immunizations within 1 month prior to screening visit.

Sex/Gender

• Sexes Eligible for Study: All

• Ages: 18 Years to 50 Years (Adult)
• Accepts Healthy Volunteers: Yes
• Contacts: Contact information is only displayed when the study is recruiting subjects
• Listed Location Countries: United States

Administrative Information

• NCT Number: NCT01489956
• Other Study ID Numbers: DAIT ITN047AI
• Has Data Monitoring Committee: Yes
• U.S. FDA-regulated Product: Not Provided
• IPD Sharing Statement: Not Provided
• Responsible Party: National Institute of Allergy and Infectious Diseases (NIAID)
• Study Sponsor: National Institute of Allergy and Infectious Diseases (NIAID)
• Collaborators: Immune Tolerance Network (ITN)

Investigators

• Study Chair: Lloyd Mayer, MD; Icahn School of Medicine at Mount Sinai

• PRS Account: National Institute of Allergy and Infectious Diseases (NIAID)
• Verification Date: March 2016