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Completed

NCT Number: NCT01910662

Characterisation of T-cell Response to Keyhole Limpet Hemocyanin (KLH) and Tuberculin Purified Protein Derivative (PPD)

This is an exploratory study to develop methodologies for the assessment of T-cell mediated therapies via skin immune challenges studies in healthy volunteers. The study will investigate what is the most appropriate; skin challenge agent, time and methodology to sample and characterise T-cells in the delayed type hypersensitivity (DTH) skin reaction. The skin challenge agents to be used in this study will be the neoantigen Keyhole Limpet Hemocyanin (KLH) and recall antigen Tuberculin Purified Protein Derivative (PPD). Part A of the study will assess an intradermal (ID) KLH challenge in three subjects to assess if the immune response to KLH is initiated by the innate or adaptive immune system. Each subject will receive one ID dose of 0.1 milligram (mg) KLH and will be assessed for a skin inflammatory response. Part B of the study will assess repeat ID challenges 28 days apart; the objective will be to characterise the T-cell response to each challenge and the kinetics of that response.

For Cohorts 1A and 1B, 16 subjects will receive an initial subcutaneous (SC) 5 mg dose of KLH. Fifteen days later the subjects will receive 0.1 mg ID KLH dose and the response will then be assessed 48 and 120 hours after the initial challenge. The ID KLH challenge will then be repeated 28 days later and the response will be assessed at 48 hours (Cohort 1A, 8 subjects) and at 120 hours (Cohort 1B, 8 subjects) post challenge. For Cohort 2, a repeat challenge of either 2 tuberculin Unit (TU) or 10 TU ID PPD will be administered 28 days apart, to 8 subjects. The first challenge response will be assessed 48 and 120 hours post challenge. The second challenge will be administered 28 days after the first and will be assessed at the same timepoints. The repeat challenge 28 days later will allow an intra-subject analysis and will determine if a subject can be used as their own control. Part C of the study will assess repeat ID challenges of PPD and PBS; the objective will be to characterise the T-cell response to each challenge and the kinetics of that response. For Cohort 1, 6 subjects will receive a 0.1ml ID dose of PBS and another dose 24 hours later. Each challenge will be assessed 48 hours post challenge. Approximately 2 subjects will have an assessment of their normal skin as a control. For Cohort 2, a repeat challenge of either 2TU or 10TU ID PPD will be administered 28 days apart, to 6 subjects. The first challenge response will be assessed 48 post challenge. The second challenge will be administered 28 days after the 1st and will be assessed at the same timepoint. The repeat challenge 28 days later will allow an intra-subject analysis and will determine if a subject can be used as their own control.

The total duration of this study for Part A is 14-18 days plus up to 30 days screening. The total duration of this study for Cohort 1A is 56-60 days plus up to 30 days screening. The total duration of this study for Cohort 1B is 59-63 days plus up to 30 days screening. The total duration of this study for Cohort 2 if 2TU is used is 45-49 days plus up to 30 day screening. The total duration of this study for Cohort 2 if 10TU is used is 48-51 days plus up to 30 days screening. The total duration of this study for Part C Cohort 1 is 15-19 days plus up to 30 day screening. The total duration of this study for Cohort 2 if 2TU is used is 42-46 days plus up to 30 day screening. The total duration of this study for Cohort 2 if 10TU is used is 45-49 days plus up to 30 days screening.

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Key information

Conditions

Dermatitis Erythema Hypersensitivity Hypersensitivity, Delayed Immune System Diseases Pathological Conditions, Signs and Symptoms Signs and Symptoms Skin Diseases Skin Manifestations Skin and Connective Tissue Diseases

Age range

18 year–55 year

Sex eligibility

All sexes

Study type

Interventional

Phase

Phase 1

Primary location

GSK Investigational Site

Cambridge, CB2 2GG, United Kingdom

Who can participate

Healthy volunteers accepted: Yes

Only the study team can determine whether someone qualifies for participation.

Inclusion criteria

  • Male or female between 18 and 55 years of age inclusive, at the time of signing the informed consent.
  • Healthy as determined by a responsible and experienced physician, based on a medical Evaluation.
  • Body weight >= 50 kg and body mass index (BMI) within the range 19.5 to 29.9 kilogram/meter squared (inclusive).
  • Female subjects of non-child bearing potential. Females of child bearing potential are eligible to enter if they are not pregnant and willing to use protocol-specified methods of contraception to prevent pregnancy.
  • Male subjects with female partners of child-bearing potential must agree to use one of the contraception methods from the time of the first dose of challenge agent until follow-up.
  • Capable of giving written informed consent.
  • Alanine aminotransferase (ALT) alkaline phosphatase and bilirubin <or=1.5x upper limit of normal (ULN).
  • Normal electrocardiogram (ECG) measurements. Average QT duration corrected for heart rate by Fridericia's formula (QTcF) <450 millisecond (msec).
  • Part B and C cohort 2 only: Subjects with a history of Bacillus Calmette Guérin (BCG) vaccination as evidence by either a BCG scar and verbal confirmation of BCG vaccination or documented medical history of a BCG vaccination with or without a BCG scar

Exclusion criteria

  • Prior medical history of anaphylaxis, severe adverse reaction to vaccines, allergy to shellfish, asthma (excluding childhood asthma), allergic rhinitis or atopic dermatitis, severe adverse reaction to local anaesthetic, previous tuberculosis infection.
  • Current treatment with beta-blockers or angiotensin converting enzyme (ACE) inhibitors.
  • History of sensitivity to any of the study challenge agents or components thereof or a history of drug or other allergy that, in the opinion of the investigator or GlaxoSmithKline (GSK) Medical Monitor, contraindicates their participation.
  • Received live, attenuated or recombinant vaccine(s) within 2 months of the first KLH or PPD injection or will require vaccination prior to the follow-up visit.
  • Part B and C cohort 2 only: From a high risk area of the world for tuberculosis or have close family members with confirmed Mycobacterium tuberculosis (MTB).
  • Antibiotics or antiviral therapy after a serious illness within 30 days of study entry.
  • Immunodeficiency or autoimmunity, assessed by medical history.
  • Presence of tattoos, naevi or other skin abnormalities such as keloids (or history of keloids) that may, in the opinion of the investigator, interfere with study assessments.
  • Use of nicotine patches on the arm at screening that would interfere with the injection sites.
  • Subjects participating, within 7 days of screening, in recreational sun-bathing, or use of sun-bed, on the area of the skin from wrist to shoulder inclusive.
  • The subject has a phobia to needles or minor surgical procedures.
  • Current or chronic history of liver disease, or known hepatic or biliary abnormalities (with the exception of Gilbert's syndrome or asymptomatic gallstones).
  • History of regular alcohol consumption within 6 months of the study.
  • Use of prescription drugs or non-prescription drugs, including NSAIDs, if in the opinion of the Investigator the medication will interfere with the study procedures or compromise subject safety. Subjects must not currently take any of the following: topical steroid cream on the arms, oral or systemic steroids or any other immune-modulators.
  • A positive pre-study Hepatitis B surface antigen or positive Hepatitis C antibody result within 3 months of screening
  • A positive pre-study drug/alcohol screen.
  • A positive test for human immunodeficiency virus (HIV) antibody.
  • Pregnant or lactating females.
  • Donation of more than 500 milliliter (mL) blood within a 56 day period.
  • Participated in a clinical trial and has received an investigational product within the following time period prior to the first dosing day in the current study: 30 days, 5 half-lives or twice the duration of the biological effect of the investigational product (whichever is longer)
  • Exposure to more than four new chemical entities within 12 months prior to the first dosing day.
  • Part A and part B Cohort 1: The subject has received KLH SC or ID in the past.

Treatment and study plan

KLH 0.1 mg

Other

KLH will be administered as single ID injection of 0.1 mg as 0.1mL of KLH (1 mg/mL)

KLH 5 mg

Other

KLH will be administered as single SC injection of 5 mg

PBS (0.1 mL)

Other

PBS will be administered in Part A on Day 1 as 3 ID injections and in Part C: Cohort 1 on Day 1 and Day 2 as a single ID injection

Tuberculin PPD

Other

Tuberculin PPD will be administered in Part B Cohort 2 on Day 1 as single ID injection of 2TU (0.04 microg/mL) in each forearm. In Part C Cohort 2 PPD will be administered on Day 1 as a single ID injection of 2TU. On day 3, if the induration is 6 mm or more; 2 ID injections of 2 TU will be administered on Day 29 in Part B Cohort 2 and 1injection of 2TU on Day 29 in Part C Cohort 2. On day 3, if the induration is less than 6 mm, subject will be re-challenged with 10 TU (0.04 microg/mL) PPD followed by 2 ID injections of 10 TU on Day 29 in Part B Cohort 2 or 1 ID injection of 10TU on Day 29 in Part C Cohort 2.

Primary outcomes

  1. Part A: Skin inflammatory response measurement as induration by the ball point pen technique

    Time frame: Over a four day period after challenge

    The induration of wheals (defined as elevation and induration that are palpable in the skin) that develop as a result of intradermal immunisation with KLH or PPD will be measured in the vertical and horizontal plane using the ball point pen technique. The diameter of the induration will be measured by rolling a medium ball point pen from 10 to 20 millimetre (mm) outside the edge of the induration and toward the centre with just enough pressure to indent the skin slightly.

  2. Part A: Skin inflammatory response measurement as erythema diameter by Laser Doppler Imaging (LDI) and ruler measurement

    Time frame: Over a four day period after challenge

    A Laser Doppler Imager will measure the area occupied by a given wheal and associated erythema, which reflects superficial blood flow of a subject's arm. Erythema at the immunisation site will be measured in the vertical and horizontal plane using a plastic flexible mm ruler. Measurements will be taken for both challenge agent and buffer control injections. Place the zero line of the ruler just inside the left edge of the area of erythema and read the ruler line just inside the right edge.

  3. Part A: Skin inflammatory response measurement as induration by characterisation and measurement of the T-cells in skin biopsies

    Time frame: Over a three day period after challenge

    T-cells will be analysed by immunocytochemistry or other techniques to characterise the T-cells

  4. Part B: Characterisation of T-cells in a skin DTH response

    Time frame: Cohort 1: Over a five day period after first challenge. Cohort 2: Over a five day period after first challenge

    Lymphocyte Activation Gene 3 (LAG 3+) T-cells collected by Punch Biopsy or skin blister will be characterised and measured by Fluorescence activated cell sorter (FACS) analysis or other immunologic techniques

  5. Part C: Characterisation of T-cells in a skin DTH response

    Time frame: Cohort 1: Over a five day period after first challenge. Cohort 2: Over a three day period after first challenge

    Lymphocyte Activation Gene 3 (LAG 3+) T-cells collected by skin blister will be characterised and measured by Fluorescence activated cell sorter (FACS) analysis or other immunologic techniques

Secondary outcomes

  1. Part A: Characterisation and measurement of the T-cells in skin biopsies.

    Time frame: At 48hrs (Day 3) after challenge

    T-cells will be analysed by immunocytochemistry or other techniques to characterise the T-cells

  2. Part B: Skin inflammatory response measurement in Cohort 1 by the ball point pen technique

    Time frame: Cohort 1A: At 48hrs (Day 17) and 120hrs (Day 20) after first challenge. At Day 44 and 48hrs (Day 45) after second challenge. Cohort 1B: At 48hrs (Day 17) and 120hrs (Day 20) after first challenge. At Day 44, 45 and 120hrs (Day 48) after second challenge

    The induration of wheals (defined as elevation and induration that are palpable in the skin) that develop as a result of intradermal immunisation with KLH or PPD will be measured in the vertical and horizontal plane using the ball point pen technique. The diameter of the induration will be measured by rolling a medium ball point pen from 10 to 20 mm outside the edge of the induration and toward the centre with just enough pressure to indent the skin slightly.

  3. Part B: Skin inflammatory response measurement in Cohort 2 by the ball point pen technique

    Time frame: Cohort 2: At 48hrs (Day3) and 120hrs (Day 6) after first challenge. At 48hrs (Day 31) and 120hrs (Day 34) after second challenge.

    The induration of wheals (defined as elevation and induration that are palpable in the skin) that develop as a result of intradermal immunisation with KLH or PPD will be measured in the vertical and horizontal plane using the ball point pen technique. The diameter of the induration will be measured by rolling a medium ball point pen from 10 to 20 mm outside the edge of the induration and toward the centre with just enough pressure to indent the skin slightly.

  4. Part B: Erythema diameter measured in Cohort 1 by Laser Doppler Imaging (LDI) and ruler measurement.

    Time frame: Cohort 1A: At 48hrs (Day 17) and 120hrs (Day 20) after first challenge. At Day 44 and 48hrs (Day 45) after second challenge. Cohort 1B: At 48hrs (Day 17) and 120hrs (Day 20) after first challenge. At Day 44, 45 and 120hrs (Day 48) after second challenge

    A Laser Doppler Imager will measure the area occupied by a given wheal and associated erythema, which reflects superficial blood flow of a subject's arm.

  5. Part B: Erythema diameter measured in Cohort 2 by Laser Doppler Imaging (LDI) and ruler measurement.

    Time frame: Cohort 2: At 48hrs (Day3) and 120hrs (Day 6) after first challenge. At 48hrs (Day 31) and 120hrs (Day 34) after second challenge.

    A Laser Doppler Imager will measure the area occupied by a given wheal and associated erythema, which reflects superficial blood flow of a subject's arm.

  6. Part B: Characterisation and measurement of the T-cells in skin biopsies in Cohort 1.

    Time frame: Cohort 1A: At 48hrs (Day 17) and 120hrs (Day 20) after first challenge. At 48hrs (Day 45) after second challenge. Cohort 1B: At 48hrs (Day 17) and 120hrs (Day 20) after first challenge. At 120hrs (Day 48) after second challenge.

    T-cells will be analysed by immunocytochemistry or other techniques to characterise the T-cells

  7. Part B: Characterisation and measurement of the T-cells in skin biopsies in Cohort 2.

    Time frame: Cohort 2: At 48hrs (Day3) and 120hrs (Day 6) after first challenge. At 48hrs (Day 31) and 120hrs (Day 34) after second challenge.

    T-cells will be analysed by immunocytochemistry or other techniques to characterise the T-cells

  8. Part A and B: Number of subjects with adverse events as a measure of safety and tolerability

    Time frame: Part A: Day 1 to Day 4, Part B: Cohort 1A: Day 1 to Day 46.Part B: Cohort 1B:Day 1 to Day 49.Part B Cohort 2: Day 1 to 35.

    The safety and tolerability of skin challenges with KLH and PPD.administration will be assessed by adverse events (AEs), serious AE (SAE) or withdrawals due to toxicities

  9. Part A and B: changes in clinical laboratory measurements to access safety and tolerability

    Time frame: Screening and up to 14 days post last visit

  10. Part A and B: Trends in and change from baseline for blood pressure measurements

    Time frame: Screening and up to 14 days post last visit

  11. Trends in and change from baseline in measure of 12-lead ECG to access safety and tolerability

    Time frame: Screening and up to 14 days post last visit

    Electrocardiograph (ECG) is a machine that measures the electrical activity of the heart and records changes in the hearts' rhythm.

  12. Immediate local hypersensitivity assessment

    Time frame: 15-30 minutes after each SC and ID challenge immunization in each Study part/Cohort

    Examination of injection site for evidence of immediate local hypersensitivity/wheal (erythema or induration will not be taken into consideration). The diameter of the wheal (central swelling/raised skin) at immunisation site, if present at time of hypersensitivity assessment, will be measured in vertical and horizontal plane using a millimetre (mm) ruler. The average will be the diameter value. If wheal is more than 5 mm in diameter than original bleb, then subject will be labelled as having immediate local hypersensitivity (ILH) and will undergo both serum total IgE and KLH or PPD prick tes

  13. Part A and B: Trends in and change from baseline for heart rate measurements

    Time frame: Screening and up to 14 days post last visit

  14. Part C: Skin inflammatory response measurement in Cohort 2 by the ball point pen technique.

    Time frame: Cohort 2: At 48hrs (Day3) and 744hrs (Day 31).

    The induration of wheals (defined as elevation and induration that are palpable in the skin) that develop as a result of intradermal immunisation with PPD will be measured in the vertical and horizontal plane using the ball point pen technique. The diameter of the induration will be measured by rolling a medium ball point pen from 10 to 20 mm outside the edge of the induration and toward the centre with just enough pressure to indent the skin slightly.

  15. Part C: Erythema diameter measured in Cohort 2 by Laser Doppler Imaging (LDI) and ruler measurement.

    Time frame: Cohort 2: At pre-challenge (Day1) and 48hrs (Day 3) after first challenge. At pre-challenge (Day 29) and 48hrs (Day 31) after second challenge.

    A Laser Doppler Imager will measure the area occupied by a given wheal and associated erythema, which reflects superficial blood flow of a subject's arm.

  16. Part C: Characterisation and measurement of the T-cells in skin blisters in Cohort 2.

    Time frame: Cohort 2: At 48hrs (Day3) and 72hrs (Day 4) after first challenge. At 48hrs (Day 31) and 69hrs (Day 32) after second challenge.

    T-cells will be analysed by immunocytochemistry or other techniques to characterise the T-cells

  17. Part A , B and C: Number of subjects with adverse events as a measure of safety and tolerability

    Time frame: Part A: Day 1 to Day 4, Part B: Cohort 1A: Day 1 to Day 46.Part B: Cohort 1B:Day 1 to Day 49.Part B Cohort 2: Day 1 to 35. Part C: Cohort 1:Day 1 to Day 19. Part C Cohort 2: Day 1 to 46.

    The safety and tolerability of skin challenges with KLH and PPD .administration will be assessed by adverse events (AEs), serious AE (SAE) or withdrawals due to toxicities

  18. Part A, B and C: changes in clinical laboratory measurements to access safety and tolerability

    Time frame: Part A and B: Screening and up to 14 days post last visit. Part C: Screening and up to 14 days post last blister draw.

  19. Part A and B: Screening and up to 14 days post last visit. Part C: Screening and up to 14 days post last blister draw.

    Time frame: Part A and B: Screening and up to 14 days post last visit. Part C: Screening and up to 14 days post last blister draw.

  20. Trends in and change from baseline in measure of 12-lead ECG to access safety and tolerability

    Time frame: Part A and B: Screening and up to 14 days post last visit. Part C: Screening and up to 14 days post last blister draw.

    Electrocardiograph (ECG) is a machine that measures the electrical activity of the heart and records changes in the hearts' rhythm.

  21. Immediate local hypersensitivity assessments

    Time frame: Part A and B: 15-30 minutes after each SC and ID challenge immunization in each Study part/Cohort. Part C: Screening and up to 14 days post last blister draw.

    Examination of injection site for evidence of immediate local hypersensitivity/wheal (erythema or induration will not be taken into consideration). The diameter of the wheal (central swelling/raised skin) at immunisation site, if present at time of hypersensitivity assessment, will be measured in vertical and horizontal plane using a millimetre (mm) ruler. The average will be the diameter value. If wheal is more than 5 mm in diameter than original bleb, then subject will be labelled as having immediate local hypersensitivity (ILH) and will undergo both serum total IgE and KLH or PPD prick tes

  22. Part A and B: Trends in and change from baseline for heart rate measurement

    Time frame: Part A and B: Screening and up to 14 days post last visit. Part C: Screening and up to 14 days post last blister draw.

Sponsors and collaborators

Lead sponsor

GlaxoSmithKline

Industry

Registry information

Official study title

Characterisation of the T-cell Response in Delayed Type Hypersensitivity Reactions to Keyhole Limpet Hemocyanin (KLH) and Tuberculin Purified Protein Derivative (PPD) in Healthy Volunteers

Important dates

Study start
2013
Primary completion
2013
Study completion
2013
First posted
Jul 29, 2013
Registry last updated
May 15, 2017

OpenTrials presents study information sourced from ClinicalTrials.gov. The official registry record should be consulted for the latest information.

View the official ClinicalTrials.gov record (opens in a new tab)

This listing is for discovery and informational purposes only. It is not medical advice, does not guarantee that a study is recruiting, and does not determine eligibility. Contact the study team and a qualified healthcare professional when considering participation.

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